Process for total synthesis of pladienolide B and pladienolide D

ABSTRACT

[Problems to be Solved] 
     To provide an effective process for total synthesis of pladienolide B and pladienolide D having excellent anti-tumor activity and to provide useful intermediates in the above-described process.  
     [Measure for Solving the Problem] 
     A process for producing a compound represented by Formula (11):  
                 
 
     wherein P 1 , P 7 , P 8 , P 9  and R 1  are the same as defined below, characterized by including reacting a compound represented by Formula (12):  
                 
 
     wherein P 7  means a hydrogen atom or a protecting group for hydroxy group; R 1  means a hydrogen atom or a hydroxy group, with a compound represented by Formula (13):  
                 
 
     wherein P 1  means a hydrogen atom or a protecting group for hydroxy group; P 8  means a hydrogen atom, an acetyl group or a protecting group for hydroxy group; P 9  means a hydrogen atom or a protecting group for hydroxy group; or P 8  and P 9  may form together a group represented by a formula:  
                 
 
     wherein R 5  means a phenyl group which may have a substituent, in the presence of a catalyst.

TECHNICAL FIELD

The present invention relates to a process for total synthesis ofpladienolide B and pladienolide D having excellent anti-tumor activityand intermediates in the above-described process.

BACKGROUND ART

Pladienolide B represented by Formula (1′):

and Pladienolide D represented by Formula (2′):

are 12 membered-ring macrolide compounds found by Sakai et al. in aculture of Streptomyces sp. Mer-11107 strain, and are known to haveexcellent anti-tumor activity (See Patent Document 1).

However, total synthesis of pladienolide B and pladienolide D has notbeen reported at all till now, and the purification of the above culturehas been the only process for acquiring pladienolide B and pladienolideD. In addition, there was a problem that there was limitation in thestructural variety of derivatives obtained by chemical conversion frompladienolide B and pladienolide D themselves which were fermentationproducts. Furthermore, absolute configuration of pladienolide B andpladienolide D has not yet been determined (See Patent Document 1), andthe determination thereof has been a problem to be solved. Accordingly,establishment of a process for total synthesis of pladienolide B andpladienolide D has been strongly desired from the viewpoints of (1)supply of pladienolide B and pladienolide D by chemical synthesis, (2)synthesis of various pladienolide derivatives and (3) determination ofabsolute configuration of pladienolide B and pladienolide D.

In the meantime, although compounds analogous to pladienolide B andpladienolide D in structure were disclosed in Patent Documents 1 to 5,total synthesis of these compounds was not disclosed in any of thedocuments.

[Patent Document 1] WO02/60890 pamphlet

[Patent Document 2] JP-A-04-352783

[Patent Document 3] WO03/099813 pamphlet

[Patent Document 4] WO2004/011459 pamphlet

[Patent Document 5] WO2004/011611 pamphlet

DISCLOSURE OF THE INVENTION

Problems to be Solved by the Invention

An object of the present invention is to provide an effective processfor total synthesis of pladienolide B and pladienolide D havingexcellent anti-tumor activity and to provide useful intermediates in theabove-described process.

Measure for Solving the Problem

In view of the above circumstances, the present inventors have conductedintensive studies and as a result found an effective process for totalsynthesis of pladienolide B and pladienolide D and useful intermediatesin the above-described process and thus completed the present invention.

That is, the present invention provides,[1] A process for producing a compound represented by Formula (1):

wherein P¹ means a hydrogen atom or a protecting group for hydroxygroup; and R¹ means a hydrogen atom or a hydroxy group, characterized bycomprising reacting a compound represented by Formula (2):

wherein P¹ and R¹ are the same as defined above, with an acetylatingagent in the presence of a base;[2] The production process according to [1], wherein the acetylatingagent is acetic anhydride;[3] The production process according to [1] or [2], wherein the base isa combination of triethylamine with 4-dimethylaminopyridine;[4] The production process according to any one of [1] to [3] whereinthe compound represented by Formula (2) in which P¹ and R¹ are hydrogenatoms is obtained by removing protecting groups for hydroxy group at 3-and 21-positions of a compound represented by Formula (6):

wherein P² means a protecting group for hydroxy group; P³ means aprotecting group for hydroxy group; and R² means a phenyl group whichmay have a substituent, to obtain a compound represented by Formula (5):

wherein R² is the same as defined above, subsequently protecting hydroxygroups at the 3- and 21-positions of Compound (5) to obtain a compoundrepresented by Formula (4):

wherein P⁴ and P⁵ mean protecting groups for hydroxy group; and R² isthe same as defined above, subsequently removing protecting groups forhydroxy group at 6- and 7-positions of Compound (4) >—R² to obtain acompound represented by Formula (3):

wherein P⁴ and P⁵ are the same as defined above, and subsequentlyremoving protecting groups at 3- and 21-positions of Compound (3);[5] A process for producing a compound represented by Formula (6):

wherein P², P³ and R² are the same as defined below, characterized bycomprising reacting a compound represented by Formula (7):

wherein P³ means a protecting group for hydroxy group; and Het means a1-phenyl-1H-tetrazol-5-yl group, with a compound represented by Formula(8):

wherein P³ means a protecting group for hydroxy group; and R² means aphenyl group which may have substituents, in the presence of a base;[6] The production process according to [5], wherein the base ispotassium bis(trimethylsilyl)amide;[7] A process for producing a compound represented by Formula (9):

wherein P², P⁶ and R² are the same as defined below, characterized bycomprising closing a ring in a compound represented by Formula (10):

wherein P² means a protecting group for hydroxy group;R² means a phenyl group which may have substituents; andP⁶ means a protecting group for hydroxy group, in the presence of acatalyst;[8] The production process according to [7], wherein the catalyst is[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(O-isopropoxyphenylmethylene)ruthenium;[9] A process for producing a compound represented by Formula (11):

wherein P¹, P⁷, P⁸, P⁹ and R¹ are the same as defined below,characterized by comprising reacting a compound represented by Formula(12):

wherein P⁷ means a hydrogen atom or a protecting group for hydroxygroup; and R¹ means a hydrogen atom or a hydroxy group) and a compoundrepresented by Formula (13):

wherein P¹ means a hydrogen atom or a protecting group for hydroxygroup; P⁸ means a hydrogen atom, an acetyl group or a protecting groupfor hydroxy group; P⁹ means a hydrogen atom or a protecting group forhydroxy group; or P⁸ and P⁹ may form together a group represented by aformula:

wherein R⁵ means a phenyl group which may have substituents, in thepresence of a catalyst;[10] The production process according to [9], wherein the catalyst is[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(O-isopropoxyphenylmethylene)rutheniumortricyclohexylphosphine[1,3-bis(2,4,6-trimethylphenyl)-4,5-dihydroimidazol-2-ylidene][benzylidene]ruthenium(IV) dichloride;[11] A compound represented by Formula (3-1):

wherein P⁴¹ and P⁵¹ may be the same or different and mean a hydrogenatom or a protecting group for hydroxy group, provided that both of P⁴¹and P⁵¹ are or either one of P⁴¹ or P⁵¹ is a protecting group forhydroxy group, or a salt thereof;[12] A compound represented by Formula (4,5,6-1):

wherein P²⁴¹ means a hydrogen atom or a protecting group for hydroxygroup; P³⁵¹ means a hydrogen atom or a protecting group for hydroxygroup; and R²¹ means a phenyl group which may have substituents, or asalt thereof;[13] A compound represented by Formula (7-1):

wherein P³¹ means a hydrogen atom or a protecting group for hydroxygroup; and Het means a 1-phenyl-1H-tetrazol-5-yl group, or a saltthereof;[14] A compound represented by Formula (8-1):

wherein P²¹ means a hydrogen atom or a protecting group for hydroxygroup; R²¹ means a phenyl group which may have substituents; and R⁴¹means a formyl group or a group represented by —CH₂OP⁶¹, wherein P⁶¹means a hydrogen atom or a protecting group for hydroxy group, or a saltthereof;[15] A compound represented by Formula (10-1):

wherein P²¹ means a hydrogen atom or a protecting group for hydroxygroup; and P⁶¹ means a hydrogen atom or a protecting group for hydroxygroup; and R²¹ means a phenyl group which may have substituents, or asalt thereof;[16] A compound represented by Formula (12-1):

wherein P⁷¹ means a hydrogen atom or a protecting group for hydroxygroup; and R¹¹ means a hydrogen atom or a hydroxy group, or a saltthereof;[17] A compound represented by Formula (13-1):

wherein P¹¹ means a hydrogen atom or a protecting group for hydroxygroup; P⁸¹ means a hydrogen atom, an acetyl group or a protecting groupfor hydroxy group; P⁹¹ means a hydrogen atom or a protecting group forhydroxy group; or P⁸¹ and P⁹¹ may form together a group represented by aformula:

wherein R⁵¹ means a phenyl group which may have substituents, or a saltthereof;[18] A process for producing a compound represented by Formula (11′):

wherein P¹, P⁸, P⁹, R⁶, R⁷ and R⁸ are the same as defined below,characterized by comprising reacting a compound represented by Formula(12′):

wherein R⁶ means a hydrogen atom or a linear C₁₋₁₀ alkyl group which mayhave a substituent; R⁷ means a hydrogen atom or a methyl group; and R⁸means a hydrogen atom or a hydroxy group, with a compound represented byFormula (13):

wherein P¹ means a hydrogen atom or a protecting group for hydroxygroup; P⁸ means a hydrogen atom, an acetyl group or a protecting groupfor hydroxy group; and P⁹ means a hydrogen atom or a protecting groupfor hydroxy group; or P⁸ and P⁹ may form together a group represented bya formula:

wherein R⁵ means a phenyl group which may have substituents, in thepresence of a catalyst.[19] A compound represented by Formula (14):

wherein P⁷¹¹ means a hydrogen atom or a protecting group for hydroxygroup; and P¹¹¹ means hydrogen atom or hydroxy group; and R^(A) means aformyl group or a group represented by —CH₂OH, or a salt thereof.[20] A compound represented by Formula (15):

Wherein P⁷¹² means a hydrogen atom or a protecting group for hydroxygroup; and R¹¹² means a hydrogen atom or a hydroxy group, or a saltthereof.[21] A compound represented by Formula (16):

wherein P²¹¹ means a hydrogen atom or a protecting group for hydroxygroup; and P⁸¹¹ means a hydrogen atom, an acetyl group or a protectinggroup for hydroxy group; and P⁹¹¹ means a hydrogen atom or a protectinggroup for hydroxy group; and R⁴¹¹ means a formyl group or a grouprepresented by —CH₂OP⁶¹¹ (wherein P⁶¹¹ means a hydrogen atom or aprotecting group for hydroxy group), or a salt thereof.

EFFECT OF THE INVENTION

Pladienolide B and pladienolide D and various kinds of pladienolidederivatives can be effectively synthesized by using a process for totalsynthesis of pladienolide B and pladienolide D and useful intermediatesin the above-described process according to the present invention.

BEST MODE FOR CARRYING OUT THE INVENTION

Hereinbelow, meaning of terms, symbols and the like described in thisspecification are described and the present invention is described indetail.

The compounds of the present invention or salts thereof may be any ofanhydrides, hydrates or solvates.

The “acetylating agent” to be used for producing Compound (1) fromCompound (2) includes acetic anhydride and acetyl chloride, andpreferably it is acetic anhydride.

The “base” to be used for producing Compound (1) from Compound (2)includes an organic base such as triethylamine, diisopropylethylamineand pyridine, etc. or a combination of the above organic base with4-dimethylaminopyridine, and preferably it is a combination oftriethylamine with 4-dimethylaminopyridine, a combination ofdiisopropylethylamine with 4-dimethylaminopyridine or a combination ofpyridine with 4-dimethylaminopyridine, and more preferably it is acombination of triethylamine with 4-dimethylaminopyridine.

The “base” to be used for producing Compound (6) from Compound (7) andCompound (8) includes potassium bis(trimethylsilyl)amide, sodiumbis(trimethylsilyl)amide, lithium bis(trimethylsilyl)amide, etc. andpreferably it is potassium bis(trimethylsilyl)amide.

The “catalyst” to be used for producing Compound (9) by closing a ringin Compound (10) includes[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(O-isopropoxyphenylmethylene)ruthenium(following formula (a)),tricyclohexylphosphine[1,3-bis(2,4,6-trimethylphenyl)-4,5-dihydroimidazol-2-ylidene][benzylidene]ruthenium(IV) dichloride (following formula (b)),[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(2-isopropoxy-5-nitrophenylmethylene)ruthenium(following formula (c)),[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(2-isopropoxy-3-phenylphenylmethylene)ruthenium(following formula (d)),[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(2,2′-diisopropoxy-1,1′-binaphthalene-3-ylmethylene)ruthenium(following formula (e)),[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(2-methoxyphenylmethylene)ruthenium(following formula (f)),[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(2,4,5-trimethoxyphenylmethylene)ruthenium(following formula (g)),tricyclohexylphosphine[1,3-bis(2,4,6-trimethylphenyl)-2,3-dihydroimidazol-2-ylidene][benzylidene]ruthenium(IV) dichloride (following formula (h)),bistricyclohexylphosphine[3,3-diphenylprop-2-en-1-ylidene]ruthenium (IV)dichloride (following formula (i)), andbis[3-bromopyridine][1,3-bis(2,4,6-trimethylphenyl)-2-imidazolidinylidene][benzylidene]ruthenium(IV) dichloride (following formula (j)) and preferably it is[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(O-isopropoxyphenylmethylene)ruthenium(following Formula (a)).

The “catalyst” to be used for producing Compound (11) from Compound (12)and Compound (13) and for producing Compound (11′) from Compound (12′)and Compound (13) includes[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(O-isopropoxyphenylmethylene)ruthenium(following formula (a)),tricyclohexylphosphine[1,3-bis(2,4,6-trimethylphenyl)-4,5-dihydroimidazol-2-ylidene][benzylidene]ruthenium(IV) dichloride (following formula (b)),[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(2-isopropoxy-5-nitrophenylmethylene)ruthenium(following formula (c)),[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(2-isopropoxy-3-phenylphenylmethylene)ruthenium(following formula (d)),[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(2,2′-diisopropoxy-1,1′-binaphthalene-3-ylmethylene)ruthenium(following formula (e)),[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(2-methoxyphenylmethylene)ruthenium(following formula (f)),[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(2,4,5-trimethoxyphenylmethylene)ruthenium(following formula (g)),tricyclohexylphosphine[1,3-bis(2,4,6-trimethylphenyl)-2,3-dihydroimidazol-2-ylidene][benzylidene]ruthenium(IV) dichloride (following formula (h)),bistricyclohexylphosphine[3,3-diphenylprop-2-en-1-ylidene]ruthenium (IV)dichloride (following formula (i)),bis[3-bromopyridine][1,3-bis(2,4,6-trimethylphenyl)-2-imidazolidinylidene][benzylidene]ruthenium(IV) dichloride (following formula (j)),bistricyclohexylphosphine[benzylidene]ruthenium (IV) dichloride(following formula (k)), and2,6-diisopropylphenylimidoneophylidenemolybdeniumbis(hexafluoro-t-butoxide) (following formula (1)) and preferably it is[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(O-isopropoxyphenylmethylene)ruthenium(following formula (a)) andtricyclohexylphosphine[1,3-bis(2,4,6-trimethylphenyl)-4,5-dihydroimidazol-2-ylidene][benzylidene]ruthenium(IV) dichloride (following formula (b)).

The “protecting group for hydroxy group” as used in this specificationis not limited particularly as long as it is a group normally used as aprotecting group for hydroxy group in organic synthesis and specificexamples thereof include silyl type protecting groups such astert-butyldimethylsilyl group, tert-butyldiphenylsilyl group,triethylsilyl group, triisopropylsilyl group, trimethylsilyl group,diethylisopropylsilyl group, dimethylisopropylsilyl group,di-tert-butylmethylsilyl group, diphenylmethylsilyl group,trimethylsilylethoxymethyl group and trimethylsilylethyl group,alkoxyalkyl type protecting groups such as methoxymethyl group,2-methoxyethoxymethyl group, 2,2,2-trichloroethoxymethyl group,1-ethoxyethyl group, 1-methyl-1-methoxyethyl group and tetrahydropyranylgroup, benzyl type protecting groups such as benzyl group,4-methoxybenzyl group, 3,4-dimethoxybenzyl group, 2,5-dimethoxybenzylgroup, 2,3,4-trimethoxybenzyl group, 3,4,5-trimethoxybenzyl group,2-nitrobenzyl group, 4-nitrobenzyl group, 4-chlorobenzyl group,2,6-dichlorobenzyl group, 4-cyanobenzyl group, diphenylmethyl group andtriphenylmethyl group, acetyl type protecting groups such as acetylgroup, chloroacetyl group, dichloroacetyl group, trichloroacetyl group,fluoroacetyl group, difluoroacetyl group, trifluoroacetyl group,bromoacetyl group, tribromoacetyl group, methoxyacetyl group, pivaloylgroup and benzoyl group, alkoxycarbonyl type protecting groups such asmethoxycarbonyl group, 2,2,2-trichloroethoxycarbonyl group andbenzyloxycarbonyl group.

Preferable examples of “protecting group for hydroxy group” in P¹ andP¹¹ include tert-butyldimethylsilyl group, tert-butyldiphenylsilylgroup, triethylsilyl group, triisopropylsilyl group,diethylisopropylsilyl group, dimethylisopropylsilyl group, methoxymethylgroup, 1-ethoxyethyl group, tetrahydropyranyl group, benzyl group,benzoyl group, etc. and more preferably it is tert-butyldimethylsilylgroup and triethylsilyl group.

Preferable examples of “protecting group for hydroxy group” in P², P²¹,and P²¹¹ include tert-butyldimethylsilyl group, tert-butyldiphenylsilylgroup, triethylsilyl group, triisopropylsilyl group,diethylisopropylsilyl group, dimethylisopropylsilyl group, 1-ethoxyethylgroup etc. and more preferably it is tert-butyldimethylsilyl group.

Preferable examples of “protecting group for hydroxy group” in P³include tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group,triisopropylsilyl group, diethylisopropylsilyl group,dimethylisopropylsilyl group, etc. and more preferably it istert-butyldimethylsilyl group, diethylisopropylsilyl group anddimethylisopropylsilyl group, and still more preferably it isdiethylisopropylsilyl group.

Preferable examples of “protecting group for hydroxy group” in P⁴ and P⁵include chloroacetyl group, dichloroacetyl group, trichloroacetyl group,fluoroacetyl group, difluoroacetyl group, trifluoroacetyl group,bromoacetyl group, tribromoacetyl group, etc. and more preferably it isdichloroacetyl group.

Preferable examples of “protecting group for hydroxy group” in P²⁴¹include tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group,triethylsilyl group, triisopropylsilyl group, diethylisopropylsilylgroup, dimethylisopropylsilyl group, chloroacetyl group, dichloroacetylgroup, trichloroacetyl group, fluoroacetyl group, difluoroacetyl group,trifluoroacetyl group, bromoacetyl group, tribromoacetyl group, etc. andmore preferably it is tert-butyldimethylsilyl group and dichloroacetylgroup.

Preferable examples of “protecting group for hydroxy group” in P³⁵¹include tert-butyldimethylsilyl group, tert-butyldiphenylsilyl group,triisopropylsilyl group, diethylisopropylsilyl group,dimethylisopropylsilyl group, chloroacetyl group, dichloroacetyl group,trichloroacetyl group, fluoroacetyl group, difluoroacetyl group,trifluoroacetyl group, bromoacetyl group and tribromoacetyl group, andmore preferably it is tert-butyldimethylsilyl group,diethylisopropylsilyl group, dimethylisopropylsilyl group,dichloroacetyl group, etc. and still more preferably it isdiethylisopropylsilyl group and dichloroacetyl group.

Preferable examples of “protecting group for hydroxy group” in P⁶, P⁶¹and P⁶¹¹ include 4-methoxybenzyl group, 3,4-dimethoxybenzyl group,2,5-dimethoxybenzyl group, 2,3,4-trimethoxybenzyl group,3,4,5-trimethoxybenzyl group, etc. and more preferably it is4-methoxybenzyl group.

Preferable examples of “protecting group for hydroxy group” in P⁷, P⁷¹,P⁷¹¹ and P⁷¹² include tert-butyldimethylsilyl group,tert-butyldiphenylsilyl group, triisopropylsilyl group,diethylisopropylsilyl group, dimethylisopropylsilyl group, triethylsilylgroup etc. and preferably it is tert-butyldimethylsilyl group,diethylisopropylsilyl group and dimethylisopropylsilyl group,triethylsilyl group and more preferably it is diethylisopropylsilylgroup.

Preferable examples of “protecting group for hydroxy group” in P⁸, P⁸¹and P⁸¹¹ include tert-butyldimethylsilyl group, tert-butyldiphenylsilylgroup, triethylsilyl group, triisopropylsilyl group,diethylisopropylsilyl group, dimethylisopropylsilyl group, methoxymethylgroup, 1-ethoxyethyl group, tetrahydropyranyl group, benzyl group,benzoyl group, etc.

Preferable examples of “protecting group for hydroxy group” in P⁹, P⁹¹and P⁹¹¹ include methoxymethyl group, 1-ethoxyethyl group, etc.

Specific examples of “phenyl group which may have a halogen atom” in R²and R⁵ include phenyl group, 4-chlorophenyl group, 4-bromophenyl group,etc. and preferably it is phenyl group.

Specific examples of “phenyl group which may have a substituent” in R²¹and R⁵¹ include phenyl group, 4-chlorophenyl group, 4-bromophenyl group,4-nitrophenyl group, 4-methoxyphenyl group, 2,4-dimethoxyphenyl group,3,4-dimethoxyphenyl group, etc. and preferably it is phenyl group.

Specific examples of “linear C₁₋₁₀ alkyl group” in R⁶ include methylgroup, ethyl group, 1-propyl group, 1-butyl group, 1-pentyl group,1-hexyl group, 1-heptyl group, 1-octyl group, 1-nonyl group, 1-decyl,etc. and preferably it is 1-heptyl group.

Specific examples of “substituent” in R⁶ include hydroxy group which maybe protected, epoxy group, oxo group, methyl group, methoxy group,phenyl group, tetrahydrofuryl group which may have a methyl group or ahydroxy group which may be protected, etc. and preferably it is hydroxygroup, epoxy group, methyl group.

“Het” is 1-phenyl-1H-tetrazol-5-yl group specifically in the aboveformula, but besides, it may be benzothiazol-2-yl group, etc. andpreferably it is 1-phenyl-1H-tetrazol-5-yl group.

The “salt” as used in this specification is not particularly limited aslong as it can form a salt of the compound of the present invention, andfor example, it includes salts with inorganic bases and, among them,pharmacologically acceptable salts are desirable.

Preferable examples of salt with an inorganic base include alkali metalsalt such as lithium salt, sodium salt, and potassium salt, alkalineearth metal salt such as calcium salt and magnesium salt, aluminum salt,and ammonium salt.

The process for production and examples of the present invention aredescribed below in detail. The list of abbreviation used in process forproduction and examples is shown below.

<A list of abbreviation>

Bn: benzyl

Et: ethyl

DEIPS: Diethylisopropylsilyl

DME: 1,2-dimethoxyethane

DMF: N,N-dimethylformamide

Me: methyl

Ph: phenyl

PMB: p-methoxybenzyl; 4-methoxybenzyl

TBS: tert-butyldimethylsilyl

TES: triethylsilyl

THF: tetrahydrofuran

Ts: p-toluenesulfonyl

Production Process 1: Process for Producing Compound (9) UsingCyclization Reaction by Olefin Metathesis

This step is a step to produce Compound (9) by closing a ring inCompound (10) in a solvent in the presence of a catalyst in the presenceor absence of a chemical reagent.

This step can be performed by a commonly used method described in aliterature (Handbook of Metathesis Vol. 1-3, Grubbs, R. H. Ed.,WILEY-VCH, 2003), etc. More specifically, this step can be performedwith reference to the reaction conditions, work up procedures andpurification method described in [Step 8] of the Example describedbelow.

This reaction can be performed in a stream of or under atmosphere of aninert gas such as nitrogen and argon.

For Compound (10), Compound (P16) described in the Example describedbelow or a compound which can be easily produced from a commerciallyavailable compound by the method which those skilled in the art usuallyperform can be used.

As for the solvent mentioned above, there is no particular limitation aslong as it can dissolve starting materials to some extent and does notobstruct the reaction, for example, aromatic hydrocarbon solvents suchas benzene, toluene, xylene, chlorobenzene and dichlorobenzene,halogenated hydrocarbon solvents such as dichloromethane,1,2-dichloroethane chloroform and carbon tetrachloride can be used andpreferably it is an aromatic hydrocarbon solvent such as benzene,toluene, xylene, chlorobenzene and dichlorobenzene.

The above catalyst means[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(O-isopropoxyphenylmethylene)ruthenium,tricyclohexylphosphine[1,3-bis(2,4,6-trimethylphenyl)-4,5-dihydroimidazol-2-ylidene][benzylidene]ruthenium(IV) dichloride,[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(2-isopropoxy-5-nitrophenylmethylene)ruthenium,[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(2-isopropoxy-3-phenylphenylmethylene)ruthenium,[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(2,2′-diisopropoxy-1,1′-binaphthalene-3-ylmethylene)ruthenium,[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(2-methoxyphenylmethylene)ruthenium,[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(2,4,5-trimethoxyphenylmethylene)ruthenium,tricyclohexylphosphine[1,3-bis(2,4,6-trimethylphenyl)-2,3-dihydroimidazol-2-ylidene][benzylidene]ruthenium(IV) dichloride,bistricyclohexylphosphine[3,3-diphenylprop-2-en-1-ylidene]ruthenium (IV)dichloride, andbis[3-bromopyridine][1,3-bis(2,4,6-trimethylphenyl)-2-imidazolidinylidene][benzylidene]ruthenium(IV) dichloride and the like but preferably it is[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(O-isopropoxyphenylmethylene)ruthenium.

The above catalyst can be used in 0.001 to 1 time molar equivalent toCompound (10), and preferably it can be used in 0.01 to 1 molarequivalent.

The above chemical reagent means 2,6-di-tert-butyl-4-methylphenol (BHT),1,4-benzoquinone, titanium triisopropoxide, etc.

The reaction temperature usually varies depending on starting materials,solvents and the other chemical reagents used in the reaction, andpreferably it is 40° C. to reflux temperature (internal temperature ofthe reaction vessel), and more preferably it is 80° C. to refluxtemperature (internal temperature of the reaction vessel) and still morepreferably it is reflux temperature (internal temperature of thereaction vessel).

The reaction time usually varies depending on starting materials,solvents and the other chemical reagents used in the reaction andreaction temperature, and it is preferable to stir the reaction solutionat the above reaction temperature for 0.5 to 48 hours after adding thechemical reagents, and it is more preferable to stir for 1 to 8 hours,and it is still more preferable to stir for about 5 hours.Production Process 2: Process for Producing Compound (6) Using JuliaCoupling

This step is a step to produce Compound (6) by reacting Compound (7) andCompound (8) in a solvent in the presence of a base.

This step can be performed by a commonly used method described in aliterature (Blakemore, P. R.; Cole, W. J.; Kociensky, P. J.; Morley, A.,Synlett, 1998, 26-28), etc. More specifically, this step can beperformed with reference to the reaction conditions, work up proceduresand purification method described in [Step 17] of the Example describedbelow.

This reaction can be performed in a stream of or under atmosphere of aninert gas such as nitrogen and argon.

For Compound (7), Compound (P35) described in the Example describedbelow or a compound which can be easily produced from a commerciallyavailable compound by the method which those skilled in the art usuallyperform can be used.

For Compound (8), Compound (P19) described in the Example describedbelow or a compound which can be easily produced from a commerciallyavailable compound by the method which those skilled in the art usuallyperform can be used.

Compound (8) can be used in 1 to 3 times molar equivalent to Compound(7), and preferably it can be used in 1 to 2 molar equivalents, morepreferably it can be used in 1.5 molar equivalents.

As for the solvent mentioned above, there is no particular limitation aslong as it can dissolve starting materials to some extent and does notobstruct the reaction, but, for example, ether solvents such astetrahydrofuran, 1,2-dimethoxyethane and diethyl ether, aromatichydrocarbon solvents such as benzene, toluene and xylene, amide solventssuch as N,N-dimethylformamide or mixed solvents of a solvent mentionedabove with hexamethylphosphoramide (HMPA) or N,N′-dimethylpropyleneurea(DMPU) can be used and preferably it is an ether solvent such astetrahydrofuran, 1,2-dimethoxyethane and diethyl ether.

The above-described base means potassium bis(trimethylsilyl)amide,sodium bis(trimethylsilyl)amide, lithium bis(trimethylsilyl)amide,lithium diisopropylamide etc. and preferably it is potassiumbis(trimethylsilyl)amide.

The above-described base can be used in 1 to 10 times molar equivalentto Compound (7), and preferably it can be used in 1 to 3 molarequivalents, more preferably it can be used in 2 molar equivalents.

The reaction temperature usually varies depending on starting materials,solvents and the other chemical reagents used in the reaction, andpreferably it is −78° C. to −60° C. (internal temperature of thereaction vessel), and more preferably it is −78° C. (internaltemperature of the reaction vessel).

The reaction time usually varies depending on starting materials,solvents and the other chemical reagents used in the reaction andreaction temperature, and it is preferable to stir the reaction solutionat the above reaction temperature for 0.5 to 12 hours after adding thechemical reagents, and it is more preferable to stir for about 1 hour.

Production Process 3: Conversion from Compound (6) to Compound (2) (theCase where P¹ and R¹ are Hydrogen Atoms)

(a) Conversion from Compound (6) to Compound (5)

This step is a step to remove protecting groups of hydroxy group at 3-and 21-positions of Compound (6) in a solvent in the presence of achemical reagent to produce Compound (5).

This step can be performed by a commonly used method described in aliterature (Protective Groups in Organic Synthesis 3rd Edition, Green,T. W.; Wuts, P. G. M., Wiley-interscience, 1999), etc. Morespecifically, this step can be performed with reference to the reactionconditions, work up procedures and purification method described in[Step 17] of the Example described below.

As for the solvent mentioned above, there is no particular limitation aslong as it can dissolve starting materials to some extent and does notobstruct the reaction, but, for example, ether solvents such astetrahydrofuran, 1,2-dimethoxyethane and diethyl ether, alcohol solventssuch as methanol and ethanol, acetonitrile, water, etc. can be used andpreferably it is an ether solvent such as tetrahydrofuran,1,2-dimethoxyethane and diethyl ether.

As for the chemical reagent mentioned above, for example, inorganicacids such as hydrochloric acid, sulfuric acid, hydrofluoric acid andperchloric acid, organic acids such as p-toluenesulfonic acid,pyridinium p-toluenesulfonate (PPTS), trifluoromethanesulfonic acid,methanesulfonic acid, acetic acid and trifluoroacetic acid, fluoride iontype chemical reagents such as tetra-n-butylammonium fluoride, potassiumfluoride, cerium fluoride, hydrogen fluoride, hydrogen fluoride-pyridinecan be used and preferably it is a fluoride ion type chemical reagentsuch as tetra-n-butylammonium fluoride, potassium fluoride, ceriumfluoride, hydrogen fluoride and hydrogen fluoride-pyridine.

The above-described chemical reagent can be used in 2 to 10 molarequivalents to Compound (6), and preferably it can be used in 2 to 5molar equivalents.

The reaction temperature usually varies depending on starting materials,solvents and the other chemical reagents used in the reaction, andpreferably it is −20° C. to 50° C. (internal temperature of the reactionvessel), and more preferably it is 0° C. to 40° C. (internal temperatureof the reaction vessel), and still more preferably it is roomtemperature (internal temperature of the reaction vessel).

The reaction time usually varies depending on starting materials,solvents and the other chemical reagents used in the reaction andreaction temperature, and it is preferable to stir the reaction solutionat the above reaction temperature for 0.5 to 48 hours after adding thechemical reagents, and it is more preferable to stir for 0.5 to 12hours, and it is still more preferable to stir for about 1 hour.(b) Conversion from Compound (5) to Compound (4)

This step is a step to protect hydroxy groups at 3- and 21-positions ofCompound (5) in a solvent in the presence of a chemical reagent and abase to produce Compound (4).

This step can be performed by a commonly used method described in aliterature (Protective Groups in Organic Synthesis 3rd Edition, Green,T. W.; Wuts, P. G. M., Wiley-interscience, 1999), etc. Morespecifically, this step can be performed with reference to the reactionconditions, work up procedures and purification method described in[Step 18] of the Example described below.

As for the solvent mentioned above, there is no particular limitation aslong as it can dissolve starting materials to some extent and does notobstruct the reaction, but, for example, ether solvents such astetrahydrofuran, 1,2-dimethoxyethane, diethyl ether, tert-butyl methylether and cyclopentyl methyl ether, halogenated hydrocarbon solventssuch as dichloromethane, 1,2-dichloroethane and chloroform, etc. can beused and preferably it is a halogenated hydrocarbon solvent such asdichloromethane, 1,2-dichloroethane and chloroform.

As for the chemical reagent mentioned above, for example, chloroaceticanhydride, dichloroacetic anhydride, trichloroacetic anhydride,difluoroacetic anhydride, trifluoroacetic anhydride, bromoaceticanhydride, chloroacetyl chloride, dichloroacetyl chloride,trichloroacetyl chloride, fluoroacetyl chloride, difluoroacetylchloride, trifluoroacetyl chloride, bromoacetyl chloride can be used andpreferably it is dichloroacetic anhydride.

The above-described chemical reagent can be used in 2 to 10 molarequivalents to Compound (5), and preferably it can be used in 2 to 5molar equivalents.

The above-described base means an organic base such as triethylamine,diisopropylethylamine and pyridine or a combination of the above organicbase with 4-dimethylaminopyridine, and preferably it is a combination oftriethylamine, diisopropylethylamine or pyridine and4-dimethylaminopyridine, and more preferably it is a combination oftriethylamine with 4-dimethylaminopyridine.

The above-described base can be used in 2 to 10 molar equivalents toCompound (5) (when 4-dimethylaminopyridine is used,4-dimethylaminopyridine in 0.01 to 1 molar equivalent to Compound (5)),and preferably it can be used in 2 to 3 molar equivalents (0.01 to 0.1molar equivalent when 4-dimethylaminopyridine is used).

The reaction temperature usually varies depending on starting materials,solvents and the other chemical reagents used in the reaction, andpreferably it is −20° C. to −50° C. (internal temperature of thereaction vessel), and more preferably it is 0° C. to 40° C. (internaltemperature of the reaction vessel), and still more preferably it isroom temperature (internal temperature of the reaction vessel).

The reaction time usually varies depending on starting materials,solvents and the other chemical reagents used in the reaction andreaction temperature, and it is preferable to stir the reaction solutionat the above reaction temperature for 5 minutes to 24 hours after addingthe chemical reagents, and it is more preferable to stir for 0.5 to 3hours, and it is still more preferable to stir for 1 to 2 hours.(c) Conversion from Compound (4) to Compound (3)

This step is a step to remove protecting groups of hydroxy group at 6-and 7-positions of Compound (4) in a solvent in the presence of achemical reagent to produce Compound (3).

This step can be performed by a commonly used method described in aliterature (Protective Groups in Organic Synthesis 3rd Edition, Green,T. W.; Wuts, P. G. M., Wiley-interscience, 1999), etc. Morespecifically, this step can be performed with reference to the reactionconditions, work up procedures and purification method described in[Step 18] of the Example described below.

As for the solvent mentioned above, there is no particular limitation aslong as it can dissolve starting materials to some extent and does notobstruct the reaction, but, for example, alcohol solvents such asmethanol and ethanol, water, etc. can be used and preferably it is analcohol solvent such as methanol and ethanol.

As for the chemical reagent mentioned above, for example, inorganicacids such as hydrochloric acid, sulfuric acid, hydrofluoric acid andperchloric acid, organic acids such as p-toluenesulfonic acid,pyridinium p-toluenesulfonate (PPTS), trifluoromethanesulfonic acid,methanesulfonic acid, acetic acid and trifluoroacetic acid, etc. can beused and preferably it is pyridinium p-toluenesulfonate (PPTS).

The above-described chemical reagent can be used in 0.05 to 20 timesmolar equivalent to Compound (4), and preferably it can be used in 0.1to 10 molar equivalents, and more preferably it can be used in 1 to 2molar equivalents.

The reaction temperature usually varies depending on starting materials,solvents and the other chemical reagents used in the reaction, andpreferably it is 0° C. to 50° C. (internal temperature of the reactionvessel), and more preferably it is room temperature (internaltemperature of the reaction vessel).

The reaction time usually varies depending on starting materials,solvents and the other chemical reagents used in the reaction andreaction temperature, and it is preferable to stir the reaction solutionat the above reaction temperature for 0.5 to 96 hours after adding thechemical reagents, and it is more preferable to stir for about 48 hours.(d) Conversion from Compound (3) to Compound (2) (the Case where P¹ andR¹ are Hydrogen Atoms)

This step is a step to remove protecting groups of hydroxy group at 3-and 21-positions of Compound (3) in a solvent in the presence of achemical reagent to produce Compound (2) (the case where P¹ and R¹ arehydrogen atoms).

This step can be performed by a commonly used method described in aliterature (Protective Groups in Organic Synthesis 3rd Edition, Green,T. W.; Wuts, P. G. M., Wiley-interscience, 1999), etc. Morespecifically, this step can be performed with reference to the reactionconditions, work up procedures and purification method described in[Step 18] of the Example described below.

As for the solvent mentioned above, there is no particular limitation aslong as it can dissolve starting materials to some extent and does notobstruct the reaction, but, for example, alcohol solvents such asmethanol and ethanol, water, etc. can be used and preferably it is analcohol solvent such as methanol and ethanol.

As for the chemical reagent mentioned above, for example, inorganicbases such as potassium carbonate, sodium carbonate and cesiumcarbonate, organic bases such as sodium methoxide, potassium methoxide,sodium ethoxide, potassium ethoxide, sodium tert-butoxide and potassiumtert-butoxide can be used and preferably it is an inorganic base such aspotassium carbonate, sodium carbonate and cesium carbonate.

The above-described chemical reagent can be used in 0.01 to 10 molarequivalents to Compound (3), and preferably it can be used in 0.1 to 2molar equivalents, and more preferably it can be used in 1 to 1.5 molarequivalents.

The reaction temperature usually varies depending on starting materials,solvents and the other chemical reagents used in the reaction, andpreferably it is −20° C. to 70° C. (internal temperature of the reactionvessel), and more preferably it is 0° C. to 50° C. (internal temperatureof the reaction vessel), and still more preferably it is roomtemperature (internal temperature of the reaction vessel).

The reaction time usually varies depending on starting materials,solvents and the other chemical reagents used in the reaction andreaction temperature, and it is preferable to stir the reaction solutionat the above reaction temperature for 5 minutes to 24 hours after addingthe chemical reagents, and it is more preferable to stir for 10 minutesto 12 hours.Production Process 4: Process for Producing Compound (1) UsingAcetylation Reaction

This step is a step to produce Compound (1) by acetylating a hydroxygroup at 7-position of Compound (2) in a solvent in the presence of anacetylating agent and a base.

This step can be performed by a commonly used method. More specifically,this step can be performed with reference to the reaction conditions,work up procedures and purification method described in [Step 19] of theExample described below.

This reaction can be performed in a stream of or under atmosphere of aninert gas such as nitrogen and argon.

As for the solvent mentioned above, there is no particular limitation aslong as it can dissolve starting materials to some extent and does notobstruct the reaction, for example, ether solvents such astetrahydrofuran, 1,2-dimethoxyethane, diethyl ether, tert-butyl methylether and cyclopentyl methyl ether, halogenated hydrocarbon solventssuch as dichloromethane, 1,2-dichloroethane and chloroform, etc. can beused and preferably it is a halogenated hydrocarbon solvent such asdichloromethane, 1,2-dichloroethane and chloroform.

The above acetylating agent means acetic anhydride and acetyl chlorideand preferably it is acetic anhydride.

The above-described acetylating agent can be used in 1 to 5 molarequivalents to Compound (2), and preferably it can be used in 0.5 to 1.5molar equivalents, and more preferably it can be used in 0.8 to 1.2molar equivalents.

The above-described base means an organic base such as triethylamine,diisopropylethylamine and pyridine or a combination of the above organicbase with 4-dimethylaminopyridine, and preferably it is a combination oftriethylamine, diisopropylethylamine or pyridine and4-dimethylaminopyridine, and more preferably it is a combination oftriethylamine with 4-dimethylaminopyridine.

The above-described base can be used in 0.5 to 10 molar equivalents toCompound (2) (when 4-dimethylaminopyridine is used,4-dimethylaminopyridine in 0.01 to 1 molar equivalent to Compound (2)),and preferably it can be used in 0.5 to 5 molar equivalents (0.1 to 1molar equivalent when 4-dimethylaminopyridine is used), and morepreferably it can be used in 0.8 to 1.2 molar equivalents (0.1 to 0.5molar equivalent when 4-dimethylaminopyridine is used).

The reaction temperature usually varies depending on starting materials,solvents and the other chemical reagents used in the reaction, andpreferably it is −20° C. to room temperature (internal temperature ofthe reaction vessel), and more preferably it is about 0° C. (internaltemperature of the reaction vessel).

The reaction time usually varies depending on starting materials,solvents and the other chemical reagents used in the reaction andreaction temperature, and it is preferable to stir the reaction solutionat the above reaction temperature 0.5 to 24 hours after adding thechemical reagents, and it is more preferable to stir for 0.5 to 8 hours,and it is still more preferable to stir for about 1 hour.Production Process 5: Process for Producing Compound (11) and Compound(11′) Using Cross Coupling Reaction by Olefin Metathesis

This step is a step to produce Compound (11) or Compound (11′) byreacting Compound (12) or Compound (12′) with Compound (13) in a solventin the presence of a catalyst.

This step can be performed by a commonly used method described in aliterature (Handbook of Metathesis Vol. 1-3, Grubbs, R. H. Ed.,WILEY-VCH, 2003), etc. More specifically, this step can be performedwith reference to the reaction conditions, work up procedures andpurification method described in [Step D6] of the Example describedbelow.

This reaction can be performed in a stream of or under atmosphere of aninert gas such as nitrogen and argon.

For Compound (12), Compound (Q9) described in the Example describedbelow or a compound which can be easily produced from a commerciallyavailable compound by the method which those skilled in the art usuallyperform can be used.

For Compound (12′), Compound (Q14) described in the Example describedbelow or a compound which can be easily produced from a commerciallyavailable compound by the method which those skilled in the art usuallyperform can be used.

For Compound (13), Compound (Q12) described in the Example describedbelow or a compound which can be easily produced from a commerciallyavailable compound by the method which those skilled in the art usuallyperform can be used.

Compound (12) or Compound (12′) can be used in 1 to 10 molar equivalentsto Compound (13), and preferably it can be used in 1.5 to 4 molarequivalents, and more preferably it can be used in 2 to 3 molarequivalents.

As for the solvent mentioned above, there is no particular limitation aslong as it can dissolve starting materials to some extent and does notobstruct the reaction, but, for example, aromatic hydrocarbon solventssuch as benzene, toluene, xylene, chlorobenzene and dichlorobenzene,halogenated hydrocarbon solvents such as dichloromethane,1,2-dichloroethane, chloroform and carbon tetrachloride, water,methanol, etc. can be used and preferably it is a halogenatedhydrocarbon solvents such as dichloromethane, 1,2-dichloroethane,chloroform and carbon tetrachloride.

The above catalyst means[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(O-isopropoxyphenylmethylene)ruthenium,tricyclohexylphosphine[1,3-bis(2,4,6-trimethylphenyl)-4,5-dihydroimidazol-2-ylidene][benzylidene]ruthenium(IV) dichloride,[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(2-isopropoxy-5-nitrophenylmethylene)ruthenium,[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(2-isopropoxy-3-phenylphenylmethylene)ruthenium,[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(2,2′-diisopropoxy-1,1′-binaphthalene-3-ylmethylene)ruthenium,[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(2-methoxyphenylmethylene)ruthenium,[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(2,4,5-trimethoxyphenylmethylene)ruthenium,tricyclohexylphosphine[1,3-bis(2,4,6-trimethylphenyl)-2,3-dihydroimidazol-2-ylidene][benzylidene]ruthenium(IV) dichloride,bistricyclohexylphosphine[3,3-diphenylprop-2-en-1-ylidene]ruthenium (IV)dichloride,bis[3-bromopyridine][1,3-bis(2,4,6-trimethylphenyl)-2-imidazolidinylidene][benzylidene]ruthenium(IV) dichloride, bistricyclohexylphosphine[benzylidene]ruthenium (IV)dichloride, and 2,6-diisopropylphenylimidoneophylidenemolybdeniumbis(hexafluoro-t-butoxide) and the like but preferably it is[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(O-isopropoxyphenylmethylene)rutheniumandtricyclohexylphosphine[1,3-bis(2,4,6-trimethylphenyl)-4,5-dihydroimidazol-2-ylidene][benzylidene]ruthenium(IV) dichloride.

The above catalyst can be used in 0.001 to 1 molar equivalent toCompound (13), and preferably it can be used in 0.01 to 0.3 molarequivalent.

The reaction temperature usually varies depending on starting materials,solvents and the other chemical reagents used in the reaction, andpreferably it is 20° C. to refluxing temperature (internal temperatureof the reaction vessel), and more preferably it is refluxing temperature(internal temperature of the reaction vessel).

The reaction time usually varies depending on starting materials,solvents and the other chemical reagents used in the reaction andreaction temperature, and it is preferable to stir the reaction solutionat the above reaction temperature for 0.1 to 96 hours after adding thechemical reagents, and it is more preferable to stir for 0.5 to 12hours, and it is still more preferable to stir for about 1 hour.

After the reaction of each of the above steps is completed, objectcompounds in each of the above steps can be taken from the reactionmixture by an usual manner.

EXAMPLES

Hereinbelow, Examples are described in order to facilitate the presentinvention to be understood, but, needless to say, the present inventionis not limited to these.

[Determination of Absolute Configuration of Pladienolide B]

Test Example 1 Determination of Relative Configuration of Pladienolide Bby NMR

[Method]

NMR information (¹H-¹H coupling constant, ¹H-¹³C long range couplingconstant and NOE correlation) which was necessary for determiningrelative configuration was obtained under the following conditions.

Device used: Unity INOVA 500 MHz (product of Varian) and JNMα 600 MHz(product of JEOL)

Solvent: deuterated pyridine

Measurement temperature: 30° C.

Sample density: about 20 mg/mL

Measuring method: ¹H-NMR, ¹³C-NMR, COSY, NOESY, HMQC, HMBC, HETLOC,J-resolved-HMBC, ¹H-decoupling

[Results]

NMR information shown in the following Table 1 to Table 3 was obtained.The relative configuration of five asymmetric carbons present inmacrolactone ring moiety was determined to be (3R*, 6R*, 7S*, 10S*,11S*) from ¹H-¹H coupling constant and NOE correlation. As for relativeconfiguration of five asymmetric carbons present in side-chain moieties,analysis by J-based configuration analysis (N. Matsumori et al, J. Org.Chem., 64, 866-876, 1999) was performed based on ¹H-¹H couplingconstant, ¹H-¹³C long range coupling constant and NOE correlation, andwas determined to be (16S*, 18R*, 19R*, 20R*, 21S*).

TABLE 1 J_(HH) multiplicity, long range J_(CH), position δ_(C) δ_(H) J =Hz J = Hz NOESY*  1 170.9 — — — —  2α 40.2 2.7  dd, 14.0, 3.2 — H2β(s),H3(m), H4α(n), H4β(n) H5α(n), H5β(w),  2β 2.78 dd, 14.0, 4.0 — H2α(s),H3(w), H4α(n), H4β(n), H5α(n), H5β(n),  3 69.8 4.1  dddd, 10.0, 4.0,²J_(H2β/C3) = 4, H2α(m), H2β(w), H4α(w), 3.5, 3.5, 3.2 ²J_(H2α/C3) < 2,H4β(n), H5α(w), H5β(n)  4α 30.8 1.95 dddd, 13.5, 13.2, — H2α(n), H2β(n),H3(w), 3.5, 3.5 H4β(s), H5α(m), H5β(o), H7(n), H8(n), H9(n), 6-  4β 1.82dddd, 13.5, 13.0, — H2α(n), H2β(n), H3(n), 10.0, 3.7 H4α(s), H5α(o),H5β(w), H7(s), H8(n), H9(w), 6-CH₃(n)  5α 37.7 1.71 ddd, 13.2, 13.0, —H2α(n), H2β(n), H3(w), 3.5 H4α(m), H4β(o), H5β(s), H7(n), H8(n), H9(n),6-CH₃(s)  5β 2.11 ddd, 13.2, 13.2, — H2α(w), H2β(n), H3(n), 3.7 H4α(o),H4β(w), H5β(s), H7(n), H8(w), H9(n), 6-CH₃(n)  6 73 — ³J_(H8/C6) < 2 — 7 79.7 5.54 d, 9.8 ³J_(H9/C7) = 6.5 H4α(n), H4β(s), H5α(n), H5β(n),H8(w), H9(w), H10(n), 6-CH₃(m),7-  8 127.4 6.27 dd, 15.2, 9.8 ²J_(H7/C8)= 3 H4α(n), H4β(n), H5α(n), H5β(w), H7(w), H9(w), H10(n), H11(w),6-CH₃(n), 7- COCH₃(n), 10-CH₃(m)  9 140.1 5.81 dd, 15.2, 10.0³J_(H11/C9) = 2.5 H4α(n), H4β(w), H5α(n), H5β(n), H7(w), H8(w), H10(n),H11(w), 6-CH₃(n), 7- COCH₃(n), 10-CH₃(m) 10 40.9 2.67 ddq, 10.7, 10.0,³J_(H8/C10) = 6, H7(n), H8(w), H9(n), H11(w), 6.8 ²J_(H9/C10) = 3,6-CH₃(n), 7-COCH₃(n), 10- ²J_(H11/C10) = 3 CH₃(s), 12-CH₃(s) 11 82.95.33 d, 10.7 ³J_(H9/C11) = 2.5, H2α(n), H2β(n), H7(n), H8(n),³J_(H13/C11) = 7 H9(w), H10(w), H13(w), 10- CH₃(s), 12-CH₃(w) 12 132 — —²J_(H11/C12) = 2.5, — ³J_(H14/C12) = 3

TABLE 2 J_(HH) multiplicity, long range J_(CH), position δ_(C) δ_(H) J =Hz J = Hz NOESY* 13 131.2 6.34 brd, 11.0 ³J_(H11/C13) = 5 H10(n),H11(w), H14(o), H15(w), 10-CH₃, 12-CH₃(w) 14 124.9 6.46 dd, 15.1, 11.0 —H11(n), H13(o), H15(w), H16(m), 12-CH₃(s), 16- 15 141.7 5.73 dd, 15.1,8.0 ³J_(H13/C15) = 5, H13(w), H14(w), H16(w), ³J_(H17β/C15) = 6 H17α(n),H17β(n), H18(w) ³J_(H17α/C15) = 3.5 12-CH₃(n), 16-CH₃(m) 16 35.8 2.54dddq, 8.5, 8.0, ³J_(H14/C16) = 5.5, H14(m), H15(w), H17α(n), 5.0, 6.8²J_(H17β/C16) = 4 H17β(m), H18(m), H19(n), 16- ²J_(H17α/C16) = 4.5CH₃(s) 17α 40.1 1.56 ddd, 13.9, 8.5, — H14(n), H15(n), H16(n), 5.9H17β(s), H18(w), H19(m), 16- 17β 1.7 ddd, 13.9, 5.9, — H14(n), H15(n),H16(m), 5.0 H17α(s), H18(m), H19(m), 16- CH₃(o) 18 56.9 2.88 ddd, 5.9,5.9, 2.1 ²J_(H17β/C18) = 7, H15(w), H16(m), H17α(w), ³J_(H17α/C18) = 4H17β(m), H19(w), H20(s), 16- ³J_(H20/C18) = 3.5 CH₃(w), 20-CH₃(m) 1961.9 3.02 dd, 8.1, 2.1 ³J_(H17β/C19) = 6, H15(n), H16(n), H17α(m),³J_(H17α/C19) = 2.5 H17β(m), H18(w), H20(m), ³J_(H20/C19) = 7.5, H21(m),16-CH₃(n), 20-CH₃(s) ³J_(H21/C19) = 3.5 20 42.1 1.45 ddq, 8.1, 4.0, 6.8²J_(H21/C20) < 2 H17α(n), H17β(o), H18(s), H19(m), H21(s), H22a(o),H22b(o), H23(s), 20-CH₃(s) 21 73.6 3.98 ddd, 8.3, 4.4, 4.0 ³J_(H19/C21)< 2, H18(n), H19(m), H20(s), ²J_(H20/C21) < 2 H22a(m), H22b(s), H23(s),H30(w) 22a 28.6 1.75 m ²J_(H21/C22) < 2 H19(n), H20(o), H21(m), H22b(o),H23(s), 20-CH₃(m) 22b 1.68 m H19(n), H20(o), H21(s), H22a(o), H23(s),H30(m) 23 11 1.08 t, 7.3 — H20(s), H21(s), H22a(s), H22(s), 20-CH₃(o)

TABLE 3 J_(HH) multiplicity, long range J_(CH), position δ_(C) δ_(H) J =Hz J = Hz NOESY* 6-CH₃ 24.8 1.46 s — H4α(m), H4β(n), H5α(s), H5β(n),H7(m), H8(n), 7- COCH₃(o) 7-COCH₃ 170.2 — — — — 7-COCH₃ 21.1 1.96 s —H7(w), H8(n), H9(n), 6-CH₃(o) 10-CH₃ 16.6 0.87 d, 6.8 ³J_(H9/C10-Me) <2, H8(n), H9(m), H10(s), H11(s), ³J_(H11/C10-Me) < 2 H13(w), 12-CH₃(w)12-CH₃ 12 1.8  brs ³J_(H11/C12-Me) = 4.5, H10(s), H11(w), H13(w),³J_(H13/C12-Me) = 7.5, H14(s), H15(n), H27(w), 16-CH₃ 21.2 1.09 d, 6.8 —H14(m), H15(m), H16(s), H17α(w), H17β(o), H18(w) 20-CH₃ 10.8 1.15 d, 6.8— H18(m), H19(s), H20(s), H21(w), H22a(m), H22b(m),recorded in pyridine-d5, 30° C., C₅D₄HN(H3) = 7.22 ppm, C₅D₅N(C3) =123.9 ppm*s: strong; m: moderate; w: weak; n: not detected; o: obscured byoverlapping signals

Test Example 2 MTPA Esterification of Pladienolide B

To a pyridine (3.0 ml) solution of pladienolide B (8 mg, 14.9 μmol),4-dimethylaminopyridine (1 mg, 8.1 μmol) and (S)-MTPA chloride (25 mg,99.2 μmol) were added and after allowed to stand still at roomtemperature for 12 hours, pyridine was evaporated and then water (10 ml)was added and the mixture was extracted with ethyl acetate (10 ml). Theobtained organic layer was concentrated and then purified by preparativeHPLC (J 'sphere ODS M-80, 20 mm I.D.×250 mm, acetonitrile: 0.15%trifluoroacetic acid aqueous solution=80:20→100:0, 9.0 ml/min) and roughMTPA ester fraction was concentrated to 1.5 ml. The concentratedfraction was purified by preparative HPLC (J 'sphere ODS M-80, 20 mmI.D.×250 mm, acetonitrile:water=60:40→100:0, 10.0 ml/min) and3,21-di-(R)-MTPA ester (12.0 mg, 14.0 μmol, 94.0%) of pladienolide B wasobtained.

In the same way, to a pyridine (3.0 ml) solution of pladienolide B (8mg, 14.9 μmol), 4-dimethylaminopyridine (1 mg, 8.1 μmol) and (R)-MTPAchloride (25 mg, 99.2 μmol) were added and after allowed to stand stillat room temperature for 12 hours, pyridine was evaporated and then water(10 ml) was added and the mixture was extracted with ethyl acetate (10ml). The obtained organic layer was concentrated and then purified bypreparative HPLC (J 'sphere ODS M-80, 20 mm I.D.×250 mm, acetonitrile:0.15% trifluoroacetic acid aqueous solution=80:20→100:0, 9.0 ml/min) andrough MTPA ester fraction was concentrated to 1.5 ml. The concentratedfraction was purified by preparative HPLC (J 'sphere ODS M-80, 20 mmI.D.×250 mm, acetonitrile:water=60:40→100:0, 10.0 ml/min) and3,21-di-(S)-MTPA ester (10.0 mg, 11.6 μmol, 82.9%) of pladienolide B wasobtained.

Test Example 3 Determination of Absolute Configuration of Pladienolide Bby Modified Mosher's Method

[Method]

NMR measurement of 3,21-di-(R)-MTPA ester and 3,21-di-(S)-MTPA ester ofpladienolide B was performed under the following conditions.

Device used: JNMα 600 MHz (product of JEOL)

Solvent: deuterated chloroform

Measurement temperature: 30° C.

Sample density: about 5 mg/mL

Measuring method: ¹H-NMR, COSY

[Results]

Assignment information of ¹H-NMR signals shown in the following Table 4for 3,21-di-(R)-MTPA ester and 3,21-di-(S)-MTPA ester of pladienolide Bwas obtained. As a result of having analyzed Δδ value (δ_(S)-δ_(R)) ofeach proton signal obtained from these chemical shifts, the absoluteconfiguration of 3-position asymmetric carbon was determined to be Rconfiguration, and the absolute configuration of 21-position asymmetriccarbon was determined to be S configuration. From the above results andthe results of the relative configuration determined by Test Example 1,the absolute configuration of ten asymmetric carbons present inpladienolide B was determined to be (3R, 6R, 7S, 10S, 11S, 16S, 18R,19R, 20R, 21S).

TABLE 4 S-MTPA ester R-MTPA ester

δ position (δ_(H)) (δ_(H)) (S − R)  2α 2.64 2.68 −0.04  2β 2.68 2.8−0.12  3 5.12 5.08 0.04  4α 1.71 1.64 0.07  4β 1.54 1.5 0.04  5α 1.71.46 0.24  5β 1.7 1.7 ±0.00  7 5.08 5.04 0.04  8 5.65 5.63 0.02  9 5.695.68 0.01 10 2.52 2.52 ±0.00 11 5.05 5.06 −0.01 13 6.04 6.05 −0.01 146.22 6.22 ±0.00 15 5.58 5.56 0.02 16 2.43 2.41 0.02 17α 1.46 1.4 0.0617β 1.53 1.5 0.03 18 2.66 2.63 0.03 19 2.41 2.27 0.14 20 1.54 1.48 0.0621 5.12 5.17 −0.05 22α 1.75 1.8 −0.05 22β 1.66 1.7 −0.04 23 0.83 0.91−0.08 6-CH₃ 1.22 1.17 0.05 7-COCH₃ 2.1 2.08 0.02 10-CH₃ 0.87 0.86 0.0112-CH₃ 1.71 1.72 −0.01 16-CH₃ 1.06 1.06 ±0.00 20-CH₃ 0.89 0.83 0.06recorded in chloroform-d, 30° C., CHCl₃ = 7.27 ppm[Determination of Absolute Configuration of Pladienolide D]

Test Example 4 Determination of Relative Configuration of Pladienolide Dby NMR

[Method]

NMR information (¹H-¹H coupling constant and NOE correlation) which wasnecessary for determining relative configuration was obtained under thefollowing conditions.

Device used: AVANCE 600 MHz (product of Bruker)

Solvent: deuterated pyridine

Measurement temperature: 30° C.

Sample density: about 20 mg/mL

Measuring method: ¹H-NMR, ¹³C-NMR, COSY, NOESY, HMQC, HMBC

[Results]

NMR information shown in the following Table 5 to Table 6 was obtained.The obtained ¹H-¹H coupling constant and NOE correlation pattern weresimilar to those of pladienolide B and therefore relative configurationof pladienolide D was determined to be (3R*, 6R*, 7S*, 10S*, 11S*, 16R*,18R*, 19R*, 20R*, 21S*).

TABLE 5 J_(HH) multiplicity, position C H J = Hz NOESY* 1 171.3 — — — 240.6 2.7 dd, 14.0, 3.2 H2 (s), H3 (m), H4 (n), H4 (n), H5 (n), H5 (w), 22.77 dd, 14.0, 4.2 H2 (s), H3 (n), H4 (n), H4 (n), H5 (n), H5 (n), 370.2 4.09 m H2 (m), H2 (n), H4 (w), H4 (n), H5 (w), H5 (n) 4 31.2 1.94 mH2 (n), H2 (n), H3 (w), H4 (s), H5 (w), H5 (o), H7 (n), H8 (n), H9 (n),6-CH₃ (m) 4 1.8 m H2 (n), H2 (n), H3 (o), H4 (s), H5 (o), H5 (w), H7(s), H8 (n), H9 (w), 6-CH₃ (n) 5 38.1 1.71 m H2 (n), H2 (n), H3 (w), H4(w), H4 (o), H5 (s), H7 (n), H8 (n), H9 (n), 6-CH₃ (m) 5 2.1 ddd, 13.5,H2 (w), H2 (n), H3 (n), 13.5, 3.3 H4 (o), H4 (w), H5 (s), H7 (n), H8(w), H9 (n), 6-CH₃ (n) 6 73.4 — — 7 80.2 5.53 d, 9.8 H4 (n), H4 (s), H5(n), H5 (n), H8 (w), H9 (n), H10 (n), H11 (n), 6-CH₃ (m), 7- 8 127.86.27 dd, 15.2, 9.8 H4 (n), H4 (n), H5 (n), H5 (w), H7 (w), H9 (w), H10(n), H11 (n), 6-CH₃ (n), 7- COCH₃ (n), 10-CH₃ (n) 9 140.5 5.82 dd, 15.2,H4 (n), H4 (w), H5 (n), 10.0 H5 (n), H7 (n), H8 (w), H10 (n), H11 (n),6-CH₃ (n), 7- COCH₃ (n), 10-CH₃ (m) 10 41.3 2.67 ddq, 10.7, H7 (n), H8(w), H9 (n), H11 (w), 10.0, 6.7 6-CH₃ (n), 7-COCH₃ (n), 10- CH₃ (s),12-CH₃ (s) 11 83.3 5.35 d, 10.7 H2 (n), H2 (n), H7 (n), H8 (n), H9 (n),H10 (w), H13 (w), 10- CH₃ (s), 12-CH₃ (w) 12 133.3 — — —

TABLE 6 J_(HH) multiplicity, position C H J = Hz NOESY* 13 131.4 6.45brd, 11.1 H10 (n), H11 (w), H14 (m), H15 (n), 10-CH₃ (w), 12-CH₃ (w) 14123.1 6.99 dd, 15.2, 11.1 H11 (n), H13 (m), H15 (m), 12- CH₃ (s), 16-CH₃(m), 15 144.5 6.16 d, 15.2 H13 (n), H14 (m), H17 (n), H17 (n), H18 (w),12-CH₃ (n), 16-CH₃ (m) 16 72.3 — 17 46.5 1.96 dd, 13.8, 6.5 H15 (n), H17(s), H18 (w), H19 (m), 16-CH₃ (m) 17 2.25 dd, 13.8, 5.2 H15 (n), H17(s), H18 (m), H19 (m), 16-CH₃ (m) 18 55.3 3.38 ddd, 6.5, 5.2, 2.0 H15(w), H17 (w), H17 (m), H19 (w), H20 (s), 16-CH₃ (m), 20-CH₃ (m) 19 62.23.12 dd, 7.6, 2.0 H15 (n), H17 (m), H17 (m), H18 (w), H20 (w), H21 (m),16- CH₃ (n), 20-CH₃ (s) 20 42.3 1.58 m H17 (n), H17 (o), H18 (s), H19(w), H21 (s), H22ab (o), H23 (s), 20-CH₃ (s) 21 74.1 3.99 m H18 (n), H19(m), H20 (s), H22ab (m), H23 (s), 20-CH₃ (w) 22ab 28.9 1.80-1.66 m H19(n), H20 (o), H21 (m), H23 (s), 20-CH₃ (m) 23 11.4 1.07 t, 7.4 H20 (s),H21 (s), H22ab (s), 20- CH₃ (o) 6-CH₃ 25.2 1.46 s H4 (m), H4 (n), H5(m), H5 (n), H7 (m), H8 (n), 7- COCH₃ (o) 7-COCH₃ 170.6 — — — 7-COCH ³21.5 1.96 s H7 (w), H8 (n), H9 (n), 6-CH₃ (o) 10-CH₃ 17.1 0.87 d, 6.7 H8(n), H9 (m), H10 (s), H11 (s), H13 (w), 12-CH₃ (w) 12-CH₃ 12.4 1.78 brsH10 (s), H11 (w), H13 (w), H14 (s), H15 (n), 10-CH₃ (w) 16-CH₃ 29.7 1.63s H14 (m), H15 (m), H16 (s), H17 (m), H17 (m), H18 (m) 20-CH₃ 10.6 1.17d, 7.0 H18 (m), H19 (s), H20 (s), H21 (w), H22ab (m), H23 (o)recorded in pyridine-d5, 30° C., C₅D₄HN(H3) = 7.22 ppm, C₅D₅N(C3) =123.9 ppm*s: strong; m: moderate; w: weak; n: not detected; o: obscured byoverlapping signals

Test Example 5 MTPA Esterification of Pladienolide D

A pyridine (262 μl) solution of pladienolide D (5.24 mg, 9.48 μmol) anda dichloromethane solution (2.0 ml) of 4-dimethylaminopyridine (82.2 mg,0.67 mmol) were prepared. A part (100 μl, 3.62 μmol) of the pyridinesolution of pladienolide D was taken and the dichloromethane solution(100 μl, 33.5 μmol) of 4-dimethylaminopyridine and (S)-MTPA chloride (10mg, 39.7 μmol) were added and after stirred at room temperature for 30minutes, 4-dimethylaminopyridine solution (100 μl, 33.5 μmol) and(S)-MTPA chloride (10 mg, 39.7 μmol) were further added and stirred atroom temperature for 30 minutes. Dichloromethane (2.0 ml) and water (2.0ml) were added to the reaction mixture and after stirred, thedichloromethane layer was separated and concentrated. The residue waspurified by NH-silica gel column chromatography (ethylacetate/hexane=3:1 mixed solvent), the obtained fraction of MTPA wasconcentrated by nitrogen gas spraying and then further dried underreduced pressure and 3,21-di-(R)-MTPA ester of pladienolide D wasobtained.

In the same way, a part (100 μl, 3.62 μmol) of the pyridine solution ofpladienolide D was taken and the dichloromethane solution (100 μl, 33.5μmol) of 4-dimethylaminopyridine and (R)-MTPA chloride (10 mg, 39.7μmol) were added and after stirred at room temperature for 30 minutes,4-dimethylaminopyridine solution (100 μl, 33.5 μmol) and (R)-MTPAchloride (10 mg, 39.7 μmol) were further added and stirred at roomtemperature for 30 minutes. Dichloromethane (2.0 ml) and water (2.0 ml)were added to the reaction mixture and after stirred, thedichloromethane layer was separated and concentrated. The residue waspurified by NH-silica gel column chromatography (ethylacetate/hexane=3:1 mixed solvent), the obtained fraction of MTPA wasconcentrated by nitrogen gas spraying and then further dried underreduced pressure and 3,21-di-(S)-MTPA ester of pladienolide D wasobtained.

Test Example 6 Determination of Absolute Configuration of Pladienolide Dby Modified Mosher's Method

[Method]

NMR measurement of 3,21-di-(R)-MTPA ester and 3,21-di-(S)-MTPA ester ofpladienolide D was performed under the following conditions.

Device used: INOVA 500 MHz (product of Varian)

Solvent: deuterated acetonitrile

Measurement temperature: 30° C.

Measuring method: ¹H-NMR, COSY, TOCSY, ROESY

[Results]

Assignment information of ¹H-NMR signals shown in the following Table 7for 3,21-di-(R)-MTPA ester and 3,21-di-(S)-MTPA ester of pladienolide Dwas obtained. As a result of having analyzed Δδ value (δ_(S)-δ_(R)) ofeach proton signal obtained from these chemical shifts, the absoluteconfiguration of 3-position asymmetric carbon was determined to be Rconfiguration, and the absolute configuration of 21-position asymmetriccarbon was determined to be S configuration. From the above results andthe results of the relative configuration determined by Test Example 4,the absolute configuration of ten asymmetric carbons present inpladienolide D was determined to be (3R, 6R, 7S, 10S, 11S, 16R, 18R,19R, 20R, 21S).

TABLE 7 S-MTPA ester R-MTPA ester

δ position (δ_(H)) (δ_(H)) (S − R)  2α 2.61 2.7 −0.09  2β 2.67 2.71−0.04  3 5.12 5.14 −0.02  4α 1.54 1.49 0.05  4β 1.66 1.49 0.17  5α 1.421.38 0.04  5β 1.61 1.59 0.02  7 4.99 4.93 0.06  8 5.67 5.67 ±0.00  95.52 5.51 0.01 10 2.57 2.57 ±0.00 11 4.91 4.94 −0.03 13 6.03 6.02 0.0114 6.45 6.42 0.03 15 5.81 5.73 0.08 17α 1.57 1.45 0.12 17β 1.78 1.750.03 18 2.83 2.77 0.06 19 2.41 2.14 0.27 20 1.52 1.4 0.12 21 5.08 5.11−0.03 22 1.65 1.69 −0.04 23 0.77 0.88 −0.11 6-CH₃ 1.14 1.1 0.04 7-COCH ³2.02 2.01 0.01 10-CH₃ 0.8 0.8 ±0.00 12-CH₃ 1.72 1.72 ±0.00 16-CH₃ 1.271.24 0.03 20-CH₃ 0.87 0.76 0.11recorded in acetonitrile-d3, 30° C., CHD₂CN = 1.93 ppm[Total Synthesis of Pladienolide B][Step 1]

Synthesis of (4Z)-6-[(4-methoxybenzyl)oxy]-4-methylhex-4-enal]

(1) Synthesis of1-({[(2Z)-3,7-dimethylocta-2,6-dien-1-yl]oxy}methyl)-4-methoxybenzene(P1)

Nerol (150 g, 972 mmol) was added dropwise to a DMF (1.501) suspensionof sodium hydride (60%, 38.9 g, 973 mmol) under nitrogen gas stream atroom temperature. The reaction solution was stirred at room temperaturefor one hour and then cooled to 0° C. After tetra-n-butyl ammoniumiodide (35.9 g, 97.2 mmol) was added to the reaction solution,4-methoxybenzyl chloride (148 g, 926 mmol) was added dropwise. Thereaction solution was stirred at 0° C. for one hour and then at roomtemperature for six hours. Water was added to the reaction solution,which was then extracted with n-heptane. After the obtained organiclayer was washed with water and brine (saturated solution of sodiumchloride) sequentially, it was dried over anhydrous magnesium sulfate.After the drying agent was filtered off, the organic layer wasconcentrated under reduced pressure. The residue was purified by silicagel column chromatography (Kanto Chemical, commercial name Silica gel60N, granular, neutral, 0.040 mm-0.100 mm; n-heptane:ethylacetate=1:0→30:1→25:1) to obtain the title compound (246.7 g) as acolorless oil.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 1.58 (s, 3H), 1.67 (s, 3H), 1.75 (d,J=1.2 Hz, 3H), 2.00-2.12 (m, 4H), 3.80 (s, 3H), 3.97 (dd, J=1.0, 7.0 Hz,1H), 4.43 (s, 2H), 5.04-5.12 (m, 1H), 5.40 (dt, J=1.2, 6.8 Hz, 2H),6.85-6.89 (m, 2H), 7.25-7.28 (m, 2H); 100 MHz ¹³C-NMR (CDCl₃) δ (ppm)17, 49, 23.36, 25.56, 26.59, 32.13, 55.06, 65.96, 71.62, 113.59, 121.82,123.79, 129.24, 130.55, 131.69, 140.31, 159.00; IR (neat) 2964, 2913,2855, 1612, 1512, 1247, 1093, 1067, 1037, 819 cm⁻¹; HRMS C₁₈H₂₇O₂ (M+H⁺)Calcd: 275.2011, Found: 275.2003.

(2) Synthesis of (4Z)-6-[(4-methoxybenzyl)oxy]-4-methylhex-4-enal (P2)

1-({[(2Z)-3,7-dimethylocta-2,6-dien-1-yl]oxy}methyl)-4-methoxybenzene(75.0 g, 273 mmol) was dissolved in dichloromethane (1.13 l) andpyridine (11.3 ml). This reaction solution was cooled to −78° C., towhich ozone was bubbled (flow rate 2 l/min, electric voltage 90 V) for170 minutes with stirring. After addition of dimethylsulfide (80.3 ml,1.09 mol), the reaction solution was allowed to warm to room temperatureand was stirred overnight. The reaction solution was concentrated underreduced pressure and the obtained residue was purified by silica gelcolumn chromatography (Kanto Chemical, commercial name Silica gel 60N,granular, neutral, 0.040 mm-0.100 mm; n-heptane:ethyl acetate=4:1) andthe title compound (36.9 g) was obtained as a colorless oil while1-({[(2Z)-3,7-dimethylocta-2,6-dien-1-yl]oxy}methyl)-4-methoxybenzene(14.1 g) was recovered.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 1.75 (s, 3H), 2.36 (t, J=7.6 Hz, 2H),2.47-2.55 (m, 2H), 3.81 (s, 3H), 3.98 (d, J=6.7 Hz, 2H), 4.43 (s, 2H),5.46 (brt, J=6.7 Hz, 1H), 6.86-6.90 (m, 2H), 7.25-7.27 (m, 2H), 9.75 (t,J=1.2 Hz, 1H); 100 MHz ¹³C-NMR (CDCl₃) δ (ppm) 22.99, 24.34, 42.11,55.09, 65.61, 71.82, 113.62, 122.97, 129.29, 130.25, 138.43, 159.04,201.63; IR (neat) 2935, 2835, 2727, 1722, 1612, 1514, 1249, 1072, 1035,819 cm⁻¹; HRMS C₁₅H₂₀NaO₃ (M+Na⁺) Calcd: 271.1310, Found: 271.1322

[Step 2]

Synthesis of methyl(3R,6Z)-3-{[tert-butyl(dimethyl)silyl]oxy-8-[(4-methoxybenzyl)oxy]-6-methyloct-6-enoate

(1) Synthesis of(4R)-3-{(3R,6Z)-3-hydroxy-8-[(4-methoxybenzyl)oxy-6-methyloct-6-enoyl]-4-isopropyl-1,3-oxazolidin-2-one(P3)

This step was performed with reference to the literature (Fukuzawa, S.;Matsuzawa, H.; Yoshimitsu, S.; J. Org. Chem., 2000, 65(6), 1702-1706.).(R)—N-bromoacetyl-4-isopropyl-2-oxazolidinone used in this step was alsoprepared by the method described in this literature.

Anhydrous THF distilled by using lithium aluminum hydride immediatelybefore use was used in this step.

After diiodomethane (1.20 ml, 14.9 mmol) was added to an anhydrous THF(200 ml) suspension of samarium powder (45.0 g, 299 mmol) at roomtemperature under nitrogen atmosphere, an anhydrous THF (700 ml)solution of diiodomethane (21.0 ml, 261 mmol) was added dropwise over100 minutes. After the addition was completed, the reaction solution wasstirred at room temperature for two hours and cooled to −78° C. Ananhydrous THF (280 ml) solution of(4Z)-6-[(4-methoxybenzyl)oxy]-4-methylhexa-4-enal (28.4 g, 115 mmol) and(R)—N-bromoacetyl-4-isopropyl-2-oxazolidinone (30.1 g, 120 mmol) wasadded dropwise to the reaction solution at such a rate that the reactionsolution temperature was kept below −72° C. After the reaction solutionwas stirred at −78° C. for one hour, it was warmed to room temperature.0.5N hydrochloric acid (300 ml) to the reaction solution was added toterminate the reaction. After addition of 0.5N hydrochloric acid (700ml), the reaction solution was extracted with ethyl acetate. After theobtained organic layer was washed with 5% sodium sulfite aqueoussolution, water and brine sequentially, it was dried over anhydrousmagnesium sulfate. After the drying agent was filtered off, the organiclayer was concentrated under reduced pressure. The obtained residue waspurified by silica gel column chromatography (Kanto Chemical, commercialname Silica gel 60N, granular, neutral, 0.040 mm-0.100 mm;n-heptane:ethyl acetate) to obtain the title compound (43.4 g, >98% de)as a yellow oil. Here, the optical purity was determined by HPLC using(DAICEL, commercial name CHIRALCEL OD; n-hexane:isopropylalcohol=4:1).

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.87 (d, J=7.2 Hz, 3H), 0.92 (d, J=7.2Hz, 3H), 1.56-1.64 (m, 2H), 1.76 (s, 3H), 2.08-2.18 (m, 1H), 2.30-2.40(m, 2H), 2.98 (dd, J=9.2, 16.9 Hz, 1H), 3.13 (dd, J=2.9, 16.9 Hz, 1H),3.26 (d, J=4.4 Hz, 1H), 3.80 (s, 3H), 3.93 (dd, J=7.2, 10.8 Hz, 1H),4.01-4.08 (m, 2H), 4.21 (dd, J=3.2, 9.2 Hz, 1H), 4.27 (t, J=9.2 Hz, 1H),4.40-4.49 (m, 1H), 4.44 (s, 2H), 5.46 (brt, J=6.6 Hz, 1H), 6.86-6.89 (m,2H), 7.26-7.29 (m, 2H); 100 MHz ¹³C-NMR (CDCl₃) δ (ppm) 14.55, 17.79,23.09, 27.73, 28.34, 34.38, 42.58, 55.10, 58.25, 63.37, 65.59, 66.76,71.75, 113.59, 122.02, 129.39, 130.26, 140.64, 153.95, 159.01, 172.18;IR (neat) 3455, 2963, 1781, 1699, 1514, 1388, 1303, 1248, 1207, 1061,1034 cm⁻¹; HRMS C₂₃H₃₄N₂NaO₅ (M+Na⁺) Calcd: 442.2206, Found: 442.2197;[α]_(D) ²²−60.2 (c 1.06, CHCl₃)

(2) Synthesis of methyl(3R,6Z)-3-{[tert-butyl(dimethyl)silyl]oxy}-8-[(4-methoxybenzyl)oxy]-6-methyloct-6-enoate(P4)

To a THF (175 ml) solution of(4R)-3-{(3R,6Z)-3-hydroxy-8-[(4-methoxybenzyl)oxy]-6-methyloct-6-enoyl}-4-isopropyl-1,3-oxazolidin-2-one(14.6 g, 34.8 mmol), a water (43.0 ml) solution of anhydrous lithiumhydroxide (2.50 g, 104 mmol) and 30% hydrogen peroxide solution (11.8ml) was added at 0° C. The reaction solution was stirred at roomtemperature for 13 hours. A water (20.0 ml) solution of anhydrouslithium hydroxide (3.75 g, 156 mmol) and 30% hydrogen peroxide solution(17.7 ml) was further added to the reaction solution and the reactionsolution was stirred for two hours. After sodium sulfite (21.0 g, 165mmol) was added to the reaction solution at 0° C., it was stirred atroom temperature for five minutes. Water was added to the reactionsolution and washed with chloroform. 2N hydrochloric acid was added tothe water layer to acidify, which was then extracted with ethyl acetate.The organic layer was washed with water and brine sequentially and thendried over anhydrous magnesium sulfate. After the drying agent wasfiltered off, the organic layer was concentrated under reduced pressure.

The obtained residue was dissolved in a mixed solvent of THF (220ml)-methanol (22.0 ml) and cooled to 0° C. and 2M n-hexane solution oftrimethylsilyldiazomethane (22.6 ml, 45.1 mmol) was added understirring. After the reaction solution was stirred at 0° C. for 40minutes, it was stirred at room temperature for 30 minutes. After aceticacid (1.00 ml) was added to the reaction solution and stirred at roomtemperature for 30 minutes, the reaction solution was concentrated underreduced pressure.

The obtained residue was dissolved in DMF (80.0 ml), followed byaddition of imidazole (7.09 g, 104 mmol) and tert-butyldimethylsilylchloride (7.85 g, 52.1 mmol), and the mixture was stirred at roomtemperature for 14 hours. The reaction solution was poured into water,which was then extracted with ethyl acetate. After the organic layer waswashed with water and brine sequentially, it was dried over anhydrousmagnesium sulfate. After the drying agent was filtered off, the organiclayer was concentrated under reduced pressure. The obtained residue waspurified by silica gel column chromatography (Kanto Chemical, commercialname Silica gel 60N, granular, neutral, 0.040 mm-0.100 mm;n-heptane:ethyl acetate=6:1) to obtain the title compound (12.9 g) as acolorless oil.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.03 (s, 3H), 0.06 (s, 3H), 0.87 (s, 9H),1.51-1.58 (m, 2H), 1,74 (brs, 3H), 1.96-2.05 (m, 1H), 2.08-2.18 (m, 1H),2.39 (dd, J=5.6, 14.1 Hz, 1H), 2.46 (dd, J=7.2, 14.4 Hz, 1H), 3.66 (s,3H), 3.80 (s, 3H), 3.96 (d, J=6.8 Hz, 2H), 4.08-4.16 (m, 1H), 4.42 (s,2H), 5.39 (brt, J=6.8 Hz, 1H), 6.86-6.89 (m, 2H), 7.24-7.28 (m, 2H); 100MHz ¹³C-NMR (CDCl₃) δ (ppm) −4.95, −4.63, 17.85, 23.38, 25.65, 27.50,35.97, 42.11, 51.36, 55.10, 65.85, 69.22, 71.77, 113.65, 121.78, 129.31,130.45, 140.24, 159.06, 171.94; IR (neat) 2952, 2930, 2856, 1740, 1514,1250, 1083, 1038, 836, 776 cm⁻¹; HRMS C₂₄H₄₁O₅Si (M+H⁺) Calcd: 437.2723,Found: 437.2731; [α]_(D) ²²−11.1 (c 1.04, CHCl₃)

[Step 3]

Synthesis of methyl6,7-O-benzylidene-3-O-[tert-butyl(dimethyl)silyl]-2,4,5-trideoxy-6-C-methyl-L-arabino-octonate

(1) Synthesis of methyl 3-O-[tert-butyl(dimethyl)silyl]-2,4,5-trideoxy-8-O-(4-methoxybenzyl)-6-C-methyl-L-arabino-octonate(P5)

To a mixed solution of tert-butyl alcohol (700 ml)-water (700 ml),AD-mix α (181 g) and methanesulfonamide (12.4 g, 130 mmol) were added at0° C. and the reaction solution was stirred at 0° C. for 30 minutes. Amixed solution of tert-butyl alcohol (150 ml)-water (150 ml) of methyl(3R,6Z)-3-{[tert-butyl(dimethyl)silyl]oxy}-8-[(4-methoxybenzyl)oxy]-6-methyloct-6-enoate(56.8 g, 130 mmol) was added to the reaction solution and stirred at 0°C. for 11 hours. Sodium sulfite (197 g, 1.56 mol) was added to thereaction solution and stirred at room temperature. After the reactionsolution was diluted with water, it was extracted with ethyl acetate.After the organic layer was washed with water and brine sequentially, itwas dried over anhydrous magnesium sulfate. After the drying agent wasfiltered off, the organic layer was concentrated under reduced pressure.The obtained residue was purified by silica gel column chromatography(Kanto Chemical, commercial name Silica gel 60N, granular, neutral,0.040 mm-0.100 mm; heptane:ethyl acetate=3:1) to obtain the titlecompound (56.3 g, 76% de) as a colorless oil. The optical purity wasdetermined by HPLC using a chiral column (DAICEL, commercial nameCHIRALCEL OD; n-hexane:isopropylalcohol=95:5).

400 MHz ¹H-NMR (CDCl₃) (data of the main product) δ (ppm) 0.04 (s, 3H),0.06 (s, 3H), 0.86 (s, 9H), 1.17 (s, 3H), 1.24-1.40 (m, 1H), 1.46-1.68(m, 3H), 2.39 (dd, J=5.6, 14.8 Hz, 1H), 2.45 (dd, J=6.8, 14.8 Hz, 1H),2.66 (s, 1H), 2.73 (d, J=4.8 Hz, 1H), 3.54-3.66 (m, 3H), 3.66 (s, 3H),3.81 (s, 3H), 4.09-4.18 (m, 1H), 4.46 (d, J=11.2 Hz, 1H), 4.50 (d,J=11.2 Hz, 1H), 6.87-6.91 (m, 2H), 7.23-7.27 (m, 2H); 100 MHz ¹³C-NMR(CDCl₃) δ (ppm) −4.99, −4.57, 17.85, 23.03, 25.66, 30.94, 32.86, 42.00,51.40, 55.14, 69.40, 70.81, 73.24, 73.39, 74.78, 113.81, 129.41, 129.50,159.32, 172.02; IR (neat) 3481, 2953, 2931, 1739, 1514, 1251, 1173,1084, 1038, 835, 777 cm⁻¹; HRMS C₂₄H₄₂NaO₇Si (M+Na⁺) Calcd: 493.2598,Found: 493.2556; [α]_(D) ²²−18.5 (c 2.22, CHCl₃)

(2) Synthesis of methyl6,7-O-benzylidene-3-O-[tert-butyl(dimethyl)silyl]-2,4,5-trideoxy-8-O-(4-methoxybenzyl)-6-C-methyl-L-arabino-octonate(P6)

To an anhydrous dichloromethane (90 ml) solution of methyl3-O-[tert-butyl(dimethyl)silyl]-2,4,5-trideoxy-8-O-(4-methoxybenzyl)-6-C-methyl-L-arabino-octonate(5.00 g, 10.6 mmol), benzaldehyde dimethylacetal (9.55 ml, 63.6 mmol)and pyridinium p-toluene sulfonate (133 mg, 0.53 mmol) were added. Thereaction solution was stirred at room temperature for 23 hours. Thereaction solution was poured into a saturated sodium hydrogen carbonateaqueous solution, which was then extracted with dichloromethane. Afterthe organic layer was washed with water and brine sequentially, it wasdried over anhydrous magnesium sulfate. After the drying agent wasfiltered off, the organic layer was concentrated under reduced pressure.The obtained residue was purified by silica gel column chromatography(Kanto Chemical, commercial name Silica gel 60N, granular, neutral,0.040 mm-0.100 mm; heptane:ethyl acetate=5:1→2:1) to obtain the titlecompound (5.92 g) as a colorless oil.

400 MHz ¹H-NMR (CDCl₃) (data of the main product) δ (ppm) 0.00 (s, 3H),0.01 (s, 3H), 0.85 (s, 9H), 1.19-1.38 (m, 1H), 1.35 (s, 3H), 1.56-1.73(m, 3H), 2.30 (dd, J=5.2, 14.5 Hz, 1H), 2.40 (dd, J=7.7, 14.5 Hz, 1H),3.58 (dd, J=5.0, 9.9 Hz, 1H), 3.64 (s, 3H), 3.68 (dd, J=7.1, 9.9 Hz,1H), 3.81 (s, 3H), 4.06 (dd, J=5.0, 7.1 Hz, 1H), 4.07-4.13 (m, 1H), 4.46(d, J=11.4 Hz, 1H), 4.56 (d, J=11.4 Hz, 1H), 5.88 (s, 1H), 6.86-6.90 (m,2H), 7.25-7.28 (m, 2H), 7.33-7.37 (m, 3H), 7.43-7.47 (m, 2H); 100 MHz¹³C-NMR (CDCl₃) δ (ppm) −5.04, −4.61, 17.84, 22.73, 25.65, 30.53, 31.18,42.18, 51.34, 55.12, 67.74, 69.47, 73.17, 81.23, 84.81, 102.27, 113.74,126.60, 128.14, 129.02, 129.38, 129.71, 137.84, 159.23, 172.03; IR(neat) 2953, 2930, 2857, 1739, 1514, 1250, 1092, 835, 776 cm⁻¹; HRMSC₃₁H₄₆NaO₇Si (M+Na⁺) Calcd: 581.2911, Found: 581.2905; [α]_(D) ²²+5.55(c 1.35, CHCl₃)

(3) Synthesis of methyl6,7-O-benzylidene-3-O-[tert-butyl(dimethyl)silyl]-2,4,5-trideoxy-6-C-methyl-L-arabino-octonate(P7)

To a mixed solution of dichloromethane (176 ml)-water (17.6 ml) ofmethyl6,7-O-benzylidene-3-O-[tert-butyl(dimethyl)silyl]-2,4,5-trideoxy-8-O-(4-methoxybenzyl)-6-C-methyl-L-arabino-octonate(8.80 g, 15.7 mmol), 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (5.38 g,23.7 mmol) was added at 0° C. After the reaction solution was stirred at0° C. for one hour, water (17.6 ml) was added. After the reactionsolution was stirred for two hours and a half,2,3-dichloro-5,6-dicyano-1,4-benzoquinone (2.70 g, 11.9 mmol) was added.The reaction solution was further stirred at 0° C. for three hours. Thereaction solution was poured into a sodium hydrogen carbonate aqueoussolution, which was then extracted with dichloromethane. After theorganic layer was washed with water and brine sequentially, it was driedover anhydrous magnesium sulfate. After the drying agent was filteredoff, the organic layer was concentrated under reduced pressure. Theobtained residue was purified by silica gel column chromatography (KantoChemical, commercial name Silica gel 60N, granular, neutral, 0.040mm-0.100 mm; n-heptane:ethyl acetate=5:1→4:1→2:1 to obtain the titlecompound (4.76 g) as a white solid. The title compound (4.44 g) wasobtained as a colorless crystal by recrystallization from n-hexane.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.02 (s, 3H), 0.05 (s, 3H), 0.86 (s, 9H),1.32-1.41 (m, 1H), 1.38 (s, 3H), 1.59-1.82 (m, 3H), 1.88 (dd, J=4.0, 8.4Hz, 1H), 2.38 (dd, J=5.6, 14.6 Hz, 1H), 2.46 (dd, J=7.2, 14.6 Hz, 1H),3.65 (s, 3H), 3.73 (dd, J=3.6, 8.4, 11.8 Hz, 1H), 3.84 (ddd, J=4.0, 7.6,11.8 Hz, 1H), 3.98 (dd, J=3.6, 7.6 Hz, 1H), 4.10-4.18 (m, 1H), 5.91 (s,1H), 7.36-7.40 (m, 3H), 7.45-7.49 (m, 2H); 100 MHz ¹³C-NMR (CDCl₃) δ(ppm)−4.94, −4.56, 17.91, 22.66, 25.71, 30.71, 31.38, 42.25, 51.47,61.42, 69.40, 81.33, 86.38, 102.20, 126.60, 128.36, 129.29, 137.69,172.07; IR (KBr) 3489, 2951, 2856, 1746, 1087, 1024, 836, 768, 704 cm⁻¹;HRMS C₂₃H₃₈NaO₆Si (M+Na⁺) Calcd: 461.2335, Found: 461.2368; [α]_(D)²²−4.31 (c 1.21, CHCl₃)

[Step 4]

Synthesis of(3R)-3-{[tert-butyl(dimethyl)silyl]oxy}-5-[(2S,4R,5S)-4-methyl-2-phenyl-5-vinyl-1,3-dioxolan-4-yl]pentanoicacid

(1) Synthesis of methyl(3R)-3-{[tert-butyl(dimethyl)silyl]oxy}-5-[(2S,4R,5S)-4-methyl-2-phenyl-5-vinyl-1,3-dioxolan-4-yl]pentanoate(P8)

To a dichloromethane (300 ml) solution of methyl6,7-O-benzylidene-3-O-[tert-butyl(dimethyl)silyl]-2,4,5-trideoxy-6-C-methyl-L-arabino-octonate(10.2 g, 23.3 mmol), Dess-Martin reagent (12.8 g, 30.3 mmol) was added.The reaction solution was stirred at room temperature for one hour. Thereaction solution was diluted with ether and washed with a saturatedsodium hydrogen carbonate aqueous solution containing sodium sulfite andbrine sequentially. The organic layer was dried over anhydrous magnesiumsulfate. After the drying agent was filtered off, the organic layer wasconcentrated under reduced pressure.

The obtained residue was dissolved in anhydrous THF (50.0 ml) and thissolution was added dropwise at −15° C. to an anhydrous THF (100 ml)solution of methylenetriphenylphosphorane prepared by a conventionalmethod from methyltriphenylphosphonium iodide (10.4 g, 25.7 mmol) andn-butyllithium (2.59M n-hexane solution 9.94 ml, 25.7 mmol) undernitrogen atmosphere. After the reaction solution was stirred at −15° C.for 20 minutes, it was stirred at room temperature for 30 minutes. Thereaction solution was poured into a saturated ammonium chloride aqueoussolution, which was then extracted with ethyl acetate. After the organiclayer was washed with water and brine sequentially, it was dried overanhydrous magnesium sulfate. After the drying agent was filtered off,the organic layer was concentrated under reduced pressure. The obtainedresidue was purified by silica gel column chromatography (KantoChemical, commercial name Silica gel 60N, granular, neutral, 0.040mm-0.100 mm; n-heptane:ethyl acetate=19:1→9:1) to obtain the titlecompound (7.91 g) as a colorless oil.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.00 (s, 3H), 0.02 (s, 3H), 0.85 (s, 9H),1.25-1.36 (m, 1H), 1.35 (s, 3H), 1.55-1.72 (m, 3H), 2.34 (dd, J=5.4,14.6 Hz, 1H), 2.42 (dd, J=7.4, 14.6 Hz, 1H), 3.64 (s, 3H), 4.06-4.14 (m,1H), 4.28 (ddd, J=1.2, 1.2, 6.8 Hz, 1H), 5.31 (ddd, J=1.2, 1.2, 10.6 Hz,1H), 5.42 (ddd, J=1.2, 1.2, 17.2 Hz, 1H), 5.90 (ddd, J=6.8, 10.6, 17.2Hz, 1H), 5.92 (s, 1H), 7.35-7.38 (m, 3H), 7.46-7.50 (m, 2H); 100 MHz¹³C-NMR (CDCl₃) δ (ppm) −5.05, −4.66, 17.81, 22.36, 25.63, 31.30, 31.93,42.23, 51.28, 69.50, 82.14, 87.73, 102.12, 118.69, 126.57, 128.15,128.99, 132.58, 137.90, 171.98; IR (neat) 2954, 2930, 2857, 1740, 1255,1090, 1065, 1006, 836, 776 cm⁻¹; HRMS C₂₄H₃₈NaO₅Si (M+Na⁺) Calcd:457.2386, Found: 457.2394; [α]_(D) ²²−11.5 (c 2.26, CHCl₃)

(2) Synthesis of(3R)-3-{[tert-butyl(dimethyl)silyl]oxy}-5-[(2S,4R,5S)-4-methyl-2-phenyl-5-vinyl-1,3-dioxolan-4-yl]pentanoicacid (P9)

Methyl(3R)-3-{[tert-butyl(dimethyl)silyl]oxy}-5-[(2S,4R,5S)-4-methyl-2-phenyl-5-vinyl-1,3-dioxolan-4-yl]pentanoate(14.6 g, 33.6 mmol) was dissolved in a mixed solution of THF (140ml)-methanol (140 ml)-water (70.0 ml) and anhydrous lithium hydroxide(4.02 g, 168 mmol) was added. The reaction solution was stirred at roomtemperature for four and a half hours. The reaction solution was pouredinto 0.5N hydrochloric acid, which was then extracted with ethylacetate. After the organic layer was washed with water and brinesequentially, it was dried over anhydrous magnesium sulfate. After thedrying agent was filtered off, the organic layer was concentrated underreduced pressure. The obtained residue was purified by silica gel columnchromatography (Kanto Chemical, commercial name Silica gel 60N,granular, neutral, 0.040 mm-0.100 mm; n-heptane:ethyl acetate=4:1→2:1)to obtain the title compound (14.1 g) as a colorless oil.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.05 (s, 6H), 0.87 (s, 9H), 1.26-1.34 (m,1H), 1.35 (s, 3H), 1.50-1.68 (m, 2H), 1.73-1.79 (m, 1H), 2.44 (dd,J=5.4, 15.4 Hz, 1H), 2.48 (dd, J=5.4, 15.4 Hz, 1H), 4.00-4.06 (m, 1H),4.29 (ddd, J=1.2, 1.2, 6.8 Hz, 1H), 5.32 (ddd, J=1.2, 1.2, 10.1 Hz, 1H),5.44 (ddd, J=1.2, 1.2, 17.2 Hz, 1H), 5.88 (ddd, J=6.8, 10.8, 17.2 Hz,1H), 5.93 (s, 1H), 7.35-7.38 (m, 3H), 7.47-7.49 (m, 2H); 100 MHz ¹³C-NMR(CDCl₃) δ (ppm) −5.00, −4.63, 17.88, 22.38, 25.69, 31.18, 32.03, 41.99,69.40, 82.18, 87.77, 102.17, 118.88, 126.59, 128.25, 129.10, 132.51,137.90, 177.39; IR (neat) 3036, 2956, 2930, 2886, 2857, 1712, 1254,1090, 1065, 836, 776 cm⁻³; HRMS C₂₃H₃₆NaO₅Si (M+Na⁺) Calcd: 443.2230,Found: 443.2212; [α]_(D) ²²+4.00 (c 1.09, CHCl₃)

[Step 5]

Synthesis of (2E)-4-[(4-methoxybenzyl)oxy]-2-methylbut-2-enal

(1) Synthesis of1-({[(2E)-4,4-dimethoxy-3-methylbut-2-en-1-yl]oxy}methyl)-4-methoxybenzene(P10)

Sodium hydride (60%, 407 mg, 10.2 mmol) was added to an anhydrous THF(20.0 ml) solution of triethyl phosphonoacetate (2.28 g, 10.2 mmol) at0° C. under nitrogen atmosphere. After the reaction solution was stirredat 0° C. for 15 minutes, pyruvaldehyde dimethylacetal (1.00 g, 8.47mmol) was added. After the reaction solution was stirred at 0° C. for 30minutes, it was stirred at room temperature for 10 minutes. The reactionsolution was poured into water, which was then extracted with n-hexane.After the organic layer was washed with brine, it was dried overanhydrous magnesium sulfate. After the drying agent was filtered off,the organic layer was concentrated under reduced pressure.

The obtained residue was dissolved in anhydrous THF (32.0 ml) and cooledto −78° C. under nitrogen atmosphere. Diisobutylaluminum hydride 0.95Mtoluene solution (21.3 ml, 20.3 mmol) was added dropwise to thissolution. After the reaction solution was stirred at −78° C. for 10minutes, it was stirred at room temperature for 30 minutes. After thereaction solution was ice cooled, followed by addition of 20% Rochellesalt solution, and vigorously stirred for one hour, it was extractedwith ethyl acetate. After the organic layer was washed with water andbrine sequentially, it was dried over anhydrous magnesium sulfate. Afterthe drying agent was filtered off, the organic layer was concentratedunder reduced pressure.

The obtained residue was dissolved in dimethoxyethane (12.0 ml) andcooled to 0° C. under nitrogen atmosphere. After addition of sodiumhydride (60%, 394 mg, 9.85 mmol) to this solution and stirring at 0° C.for 20 minutes, p-methoxybenzyl chloride (1.00 ml, 7.39 mmol) and sodiumiodide (1.11 g, 7.39 mmol) were added. After the reaction solution wasstirred at room temperature for 45 minutes, it was poured into water,which was then extracted with n-hexane. After the organic layer waswashed with a sodium sulfite aqueous solution and brine sequentially, itwas dried over anhydrous magnesium sulfate. After the drying agent wasfiltered off, the organic layer was concentrated under reduced pressure.The obtained residue was purified by silica gel column chromatography(Kanto Chemical, commercial name Silica gel 60N, granular, neutral,0.040 mm-0.100 mm; heptane:ethyl acetate=19:1→9:1→5:1) to obtain thetitle compound (1.52 g) as a colorless oil. The title compound wasdetermined to be a mixture of E:Z=18:7 by ¹H-NMR.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 1.62 (d, J=0.8 Hz, 2.16H), 1.74 (d, J=1.2Hz, 0.84H), 3.30 (s, 1.68H), 3.31 (s, 4.32H), 3.81 (s, 2.16H), 3.82 (s,0.84H), 4.09 (d, J=6, 0 Hz, 2H), 4.45 (s, 2H), 4.49 (s, 0.72H), 4.87 (s,0.28H), 5.66 (brt, J=6.6 Hz, 0.28H), 5.78 (brt, J=6.2 Hz, 0.72H),6.86-6.91 (m, 2H), 7.25-7.28 (m, 2H); IR (neat) 2934, 2834, 1514, 1249,1112, 1072, 1036, 820 cm⁻¹; HRMS C₁₅H₂₂AgO₄ (M+Ag⁺) Calcd: 373.0569,Found: 373.0552

(2) Synthesis of (2E)-4-[(4-methoxybenzyl)oxy]-2-methylbut-2-enal (P11)

1-({[(2E)-4,4-dimethoxy-3-methylbut-2-en-1-yl]oxy}methyl)-4-methoxybenzene(1.52 g, 5.71 mmol) was dissolved in acetonitrile (16.0 ml), followed byaddition of 1N hydrochloric acid (4.00 ml) and the reaction solution wasstirred at room temperature for one hour. Subsequently, 2N hydrochloricacid (4.00 ml) to the reaction solution was added and it was furtherstirred for four hours. The reaction solution was poured into asaturated aqueous solution of sodium hydrogen carbonate, which was thenextracted with diethyl ether. After the organic layer was washed withwater and brine sequentially, it was dried over anhydrous magnesiumsulfate. The drying agent was filtered off and the organic layer wasconcentrated under reduced pressure to obtain the title compound (1.52g) as a colorless oil.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 1.73 (dt, J=1.2, 1.2 Hz, 3H), 3.82 (s,3H), 4.32 (dq, J=1.2, 5.6 Hz, 2H), 4.52 (s, 2H), 6.60 (tq, J=1.2, 5.6Hz, 1H), 6.88-6.92 (m, 2H), 7.26-7.30 (m, 2H), 9.44 (s, 1H); 100 MHz¹³C-NMR (CDCl₃ δ (ppm) 9.20, 54.94, 66.17, 72.45, 113.63, 129.24,129.32, 139.04, 149.47, 159.18, 194.10; IR (neat) 2837, 1688, 1612,1512, 1249, 1074, 1033, 820 cm⁻¹; HRMS C₁₃H₁₆NaO₃ (M+Na⁺) Calcd:243.0997, Found: 243.1044

[Step 6]

Synthesis of(1S,3R,4S,5E)-4-{[tert-butyl(dimethyl)silyl]oxy}-7-[(4-methoxybenzyl)oxy]-1,3,5-trimethyl-2-oxohept-5-en-1-ylbenzoate

(1) Synthesis of(1S,3R,4S,5E)-4-hydroxy-7-[(4-methoxybenzyl)oxy]-1,3,5-trimethyl-2-oxohept-5-en-1-ylbenzoate (P12)

This reaction was performed with reference to the literature (Paterson,I.; Wallac, D.; Cowden, C. J.; Synthesis, 1998, 639-652.).(S)-2-benzoyloxypentane-3-one used in this step was prepared by a methoddescribed in this literature.

Anhydrous diethyl ether distilled by using lithium aluminum hydrideimmediately before use was used in this step.

N,N-dimethylethylamine (24.0 ml, 221 mmol) was added to an anhydrousdiethyl ether (500 ml) solution of dicyclohexylboron chloride (38.8 ml,177 mmol) prepared from a commercial boron monochrolide-methyl sulfidecomplex by a method described in the literature (Paterson, I.;Temal-Laib, T.; Org. Lett., 2002, 4(15), 2473-2476.) under nitrogenatmosphere −78° C. and the reaction solution was stirred at −78° C. for15 minutes. An anhydrous diethyl ether (400 ml) solution of(S)-2-benzoyloxy pentane-3-one (30.4 g, 148 mmol) was added dropwise tothis reaction solution over 35 minutes, and it was stirred at −78° C.for 10 minutes and then stirred at 0° C. for two hours. The reactionsolution was cooled to −78° C. again and an anhydrous diethyl ether (400ml) solution of (2E)-4-[(4-methoxybenzyl)oxy]-2-methylbut-2-enal (27.0g, 123 mmol) was added dropwise over 20 minutes and it was stirred at−78° C. for two hours and then stirred at −26° C. overnight. After thereaction solution was warmed to 0° C., methanol (400 ml), phosphatebuffer (pH=7, 400 ml) and 30% hydrogen peroxide solution (350 ml) wereadded sequentially and the reaction solution was stirred at 0° C. forone hour. The reaction solution was poured into water, which was thenextracted with ethyl acetate. The organic layer was washed with waterand brine sequentially and dried over anhydrous magnesium sulfate. Afterthe drying agent was filtered off, the organic layer was concentratedunder reduced pressure. The obtained residue was purified by silica gelcolumn chromatography (Kanto Chemical, commercial name Silica gel 60N,granular, neutral, 0.040 mm-0.100 mm; n-heptane:ethyl acetate=4:1→2:1).The obtained white solid was recrystallized (n-hexane:ethyl acetate) toobtain the title compound (42.6 g, >99% de) as a colorless needle. Theoptical purity was determined by HPLC using a chiral column (DAICEL,commercial name CHIRALCEL OD; n-hexane:isopropylalcohol=95:5).

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 1.07 (d, J=7.2 Hz, 3H), 1.57 (d, J=7.2Hz, 3H), 1.63 (s, 3H), 2.02 (d, J=3.8 Hz, 1H), 3.04 (dq, J=7.2, 9.2 Hz,1H), 3.81 (s, 3H), 3.80-4.10 (m, 2H), 4.24 (dd, J=3.8, 9.2 Hz, 1H), 4.44(s, 2H), 5.45 (q, J=7.2 Hz, 1H), 5.65 (brt, J=6.4 Hz, 1H), 6.86-6.90 (m,2H), 7.24-7.28 (m, 2H), 7.44-7.47 (m, 2H), 7.56-7.61 (m, 1H), 8.07-8.10(m, 2H); 100 MHz ¹³C-NMR (CDCl₃) δ (ppm) 11.07, 14.46, 15.46, 45.38,55.22, 65.75, 72.00, 75.06, 79.48, 113.75, 126.06, 128.38, 129.36,129.51, 129.74, 130.20, 133.23, 138.19, 159.18, 165.83, 210.72; IR (KBr)3466, 3050, 2991, 2967, 2932, 2871, 2836, 1731, 1719, 1513, 1453, 1301,1247, 1121, 1072, 1038, 996, 711 cm⁻¹; HRMS C₂₅H₃₀NaO₆ (M+Na⁺) Calcd:449.1940, Found: 449.1951, [α]_(D) ²⁵+30.3 (c 1.00, CHCl₃)

(2) Synthesis of(1S,3R,4S,5E)-4-{[tert-butyl(dimethyl)silyl]oxy}-7-[(4-methoxybenzyl)oxy]-1,3,5-trimethyl-2-oxohept-5-en-1-ylbenzoate (P13)

2,6-Lutidine (38.2 ml, 328 mmol) was added to an anhydrousdichloromethane (1.00 l) solution of(1S,3R,4S,5E)-4-hydroxy-7-[(4-methoxybenzyl)oxy]-1,3,5-trimethyl-2-oxohept-5-en-1-ylbenzoate (70.0 g, 164 mmol) at −78° C. under nitrogen atmosphere.Trifluoromethanesulfonic acid tert-butyldimethylsilyl ester (56.5 ml,246 mmol) was added dropwise to the reaction solution over 10 minutes.After the reaction solution was stirred at −78° C. for one and a halfhour, a saturated sodium hydrogen carbonate aqueous solution was addedat −78° C. The reaction solution was warmed to room temperature, whichwas then extracted with dichloromethane. After the organic layer waswashed with water and brine sequentially, it was dried over anhydrousmagnesium sulfate. After the drying agent was filtered off, the organiclayer was concentrated under reduced pressure. The obtained residue waspurified by silica gel column chromatography (Kanto Chemical, commercialname Silica gel 60N, granular, neutral, 0.040 mm-0.100 mm;n-heptane:ethyl acetate=8:1) to obtain the title compound (89.0 g) as acolorless oil.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) −0.02 (s, 3H), −0.01 (s, 3H), 0.82 (s,9H), 0.97 (d, J=7.2 Hz, 3H), 1.54 (d, J=7.2 Hz, 3H), 1.58 (s, 3H), 3.00(dq, J=7.2, 9.6 Hz, 1H), 3.81 (s, 3H), 4.04 (d, J=6.0 Hz, 2H), 4.28 (d,J=9.6 Hz, 1H), 4.43 (s, 2H), 5.43 (q, J=7.2 Hz, 1H), 5.59 (brt, J=6.0Hz, 1H), 6.86-6.90 (m, 2H), 7.24-7.27 (m, 2H), 7.43-7.47 (m, 2H),7.56-7.60 (m, 1H), 8.07-8.09 (m, 2H); 100 MHz ¹³C-NMR (CDCl₃) δ (ppm)−5.17, −4.78, 10.79, 14.54, 15.17, 18.01, 25.75, 46.17, 55.21, 65.83,71.72, 75.20, 80.54, 113.75, 126.24, 128.36, 129.22, 129.66, 129.76,130.35, 133.16, 138.11, 159.15, 165.68, 209.02; IR (neat) 2954, 2931,2856, 1721, 1512, 1453, 1301, 1250, 1117, 1070, 1040, 837, 778, 713cm⁻¹; HRMS C₃₁H₄₄NaO₆Si (M+Na⁺) Calcd: 563.2805, Found: 563.2786;[α]_(D) ²⁵+12.6 (c 1.01, CHCl₃)

[Step 7]

(3S,4S,5E)-7-[(4-methoxybenzyl)oxy]-3,5-dimethylhepta-1,5-dien-4-ol

(1) Synthesis oftert-butyl({(1S,2E)-4-[(4-methoxybenzyl)oxy]-2-methyl-1-[(1S)-1-methylprop-2-en-1-yl]but-2-en-1-yl}oxy)dimethylsilane(P14)

To an anhydrous THF (10.0 ml) solution of(1S,3R,4S,5E)-4-{[tert-butyl(dimethyl)silyl]oxy}-7-[(4-methoxybenzyl)oxy]-1,3,5-trimethyl-2-oxohept-5-en-1-ylbenzoate (1.00 g, 1.85 mmol), lithium boron hydride 2M THF (18.5 ml,37.0 mmol) solution was added at −78° C. under nitrogen atmosphere. Thereaction solution was stirred overnight while warmed to roomtemperature. After the reaction solution was ice cooled and water wasadded thereto, it was extracted with ethyl acetate. After the organiclayer was washed with water and brine sequentially, it was dried overanhydrous magnesium sulfate. The drying agent was filtered off and theorganic layer was concentrated under reduced pressure to obtain a whitesolid (611 mg).

300 mg from the obtained white solid was dissolved in a mixed solvent ofTHF-water (4:1, 6.00 ml). Sodium periodate (438 mg, 2.05 mmol) was addedto the reaction solution. After the reaction solution was stirred to atroom temperature for one and a half hour, it was poured into water,which was then extracted with ethyl acetate. After the organic layer waswashed with water and brine sequentially, it was dried over anhydrousmagnesium sulfate. After the drying agent was filtered off, the organiclayer was concentrated under reduced pressure.

The obtained residue was dissolved in anhydrous THF (4.00 ml) and addeddropwise at under nitrogen atmosphere at −15° C. to an anhydrous THF(4.00 ml) solution of methylenetriphenylphosphorane prepared by aconventional method from methyl iodide triphenylphosphonium (415 mg,1.02 mmol) and 2.59M n-butyllithium n-hexane (397 μl, 1.03 mmol)solution. After the reaction solution was stirred at −15° C. for onehour, it was poured into water, which was then extracted with ethylacetate. After the organic layer was washed with water and brinesequentially, it was dried over anhydrous magnesium sulfate. After thedrying agent was filtered off, the organic layer was concentrated underreduced pressure. The obtained residue was purified by silica gel columnchromatography (Kanto Chemical, commercial name Silica gel 60N,granular, neutral, 0.040 mm-0.100 mm; n-heptane:ethyl acetate=19:1) toobtain the title compound (258 mg) as a colorless oil.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) −0.03 (s, 3H), 0.02 (s, 3H), 0.87 (d,J=7.2 Hz, 3H), 0.88 (s, 9H), 1.58 (s, 3H), 2.24-2.33 (m, 1H), 3.72 (d,J=7.2 Hz, 1H), 3.81 (s, 3H), 4.05 (d, J=6.4 Hz, 2H), 4.42 (s, 2H), 4.98(brd, J=10.4 Hz, 1H), 4.99 (brd, J=17.4 Hz, 1H), 5.50 (brt, J=6.4 Hz,1H), 5.84 (ddd, J=7.6, 10.4, 17.4 Hz, 1H), 6.87-6.89 (m, 2H), 7.25-7.27(m, 2H); 100 MHz ¹³C-NMR (CDCl₃) δ (ppm) −5.02, −4.55, 11.74, 16.65,18.16, 25.81, 42.07, 55.12, 65.84, 71.32, 82.35, 113.68, 113.91, 123.76,129.19, 130.55, 140.26, 141.67, 159.07; IR (neat) 2956, 2929, 2856,1514, 1249, 1065, 1040, 836, 775 cm⁻¹; HRMS C₂₃H₃₈NaO₃Si (M+Na⁺) Calcd:413.2488, Found: 413.2504; [α]_(D) ²⁶−2.26 (c 1.02, CHCl₃)

(2) Synthesis of(3S,4S,5E)-7-[(4-methoxybenzyl)oxy]-3,5-dimethylhepta-1,5-dien-4-ol(P15)

1N hydrochloric acid (100 ml) was added to an acetonitrile (300 ml)solution oftert-butyl({(1S,2E)-4-[(4-methoxybenzyl)oxy]-2-methyl-1-[(1S)-1-methylprop-2-en-1-yl]but-2-en-1-yl}oxy)dimethylsilane(41.8 g, 107 mmol). After the reaction solution was stirred at roomtemperature for ten and a half hours, it was poured into brine andextracted with ethyl acetate. After the organic layer was washed withbrine, it was dried over anhydrous magnesium sulfate. After the dryingagent was filtered off, the organic layer was concentrated under reducedpressure. The obtained residue was purified by silica gel columnchromatography (Kanto Chemical, commercial name Silica gel 60N,granular, neutral, 0.040 mm-0.100 mm; n-heptane:ethylacetate=4:1→3:1→2:1) and was obtained title compound (26.8 g) as acolorless oil.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.93 (d, J=6.4 Hz, 3H), 1.64 (s, 3H),1.77 (brs, 1H), 2.28-2.39 (m, 1H), 3.71 (d, J=8.0 Hz, 1H), 3.81 (s, 3H),4.07 (d, J=6.4 Hz, 2H), 4.45 (s, 2H), 5.15 (brd, J=10.4 Hz, 1H), 5.16(brd, J=17.1 Hz, 1H), 5.62 (brt, J=6.4 Hz, 1H), 5.75 (ddd, J=8.4, 10.4,17.1 Hz, 1H), 6.86-6.90 (m, 2H), 7.25-7.29 (m, 2H); 100 MHz ¹³C-NMR(CDCl₃) δ (ppm) 11.44, 16.62, 41.60, 55.04, 65.74, 71.58, 80.52, 113.59,116.10, 124.73, 129.19, 130.24, 138.94, 140.67, 159.00; IR (neat) 3443,2962, 2931, 2862, 1613, 1513, 1456, 1301, 1249, 1174, 1065, 1035, 913,820 cm⁻¹; HRMS C₁₇H₂₄NaO₃ (M+Na⁺) Calcd: 299.1623, Found: 299.1612;[α]_(D) ²⁶−14.1 (c 1.02, CHCl₃)

[Step 8]

Synthesis of (2S,3aS,4E,6S,7S,13aR)-11-{[tert-butyl(dimethyl)silyl]oxy}-7-{(1E)-3-[(4-methoxybenzyl)oxy]-1-methylprop-1-en-1-yl}-6,13a-dimethyl-2-phenyl-3a,6,7,10,11,12,13,13a-octahydro-9H-[1,3]dioxolo[4,5-f]oxacyclododecin-9-one

(1) Synthesis of(1S,2E)-4-[(4-methoxybenzyl)oxy]-2-methyl-1-[(1S)-1-methylprop-2-en-1-yl]but-2-en-1-yl3-{[tert-butyl(dimethyl)silyl]oxy}-5-[(2S,4R,5S)-4-methyl-2-phenyl-5-vinyl-1,3-dioxolan-4-yl]pentanoate(P16)

This reaction was performed with reference to the literature (Inanaga,J.; Hirata, K.; Saeki, H; Katsuki, T.; Yamaguchi, M.; Bull. Chem. Soc.Jpn. 1979, 52(7), 1989-1993.).

To an anhydrous THF (130 ml) solution of(3R)-3-{[tert-butyl(dimethyl)silyl]oxy}-5-[(2S,4R,5S)-4-methyl-2-phenyl-5-vinyl-1,3-dioxolan-4-yl]pentanoicacid (6.50 g, 15.5 mmol) was added triethylamine (2.80 ml, 20.2 mmol)under nitrogen atmosphere and cooled to 0° C. 2,4,6-Trichlorobenzoylchloride (2.65 ml, 17.0 mmol) was added to the reaction solution. Afterthe reaction solution was stirred at 0° C. for 10 minutes, it wasstirred at room temperature for two hours. After the reaction solutionwas filtered with celite, the organic layer was concentrated underreduced pressure. To the obtained residue, an anhydrous toluene (130 ml)solution of(3S,4S,5E)-7-[(4-methoxybenzyl)oxy]-3,5-dimethylhepta-1,5-dien-4-ol(4.71 g, 17.0 mmol) and 4-dimethylaminopyridine (2.46 g, 20.2 mmol) wereadded under nitrogen atmosphere. After the reaction solution was stirredat room temperature for two hours, it was poured into 0.5N hydrochloricacid and extracted with ethyl acetate. After the organic layer waswashed with a saturated sodium hydrogen carbonate aqueous solution,water and brine sequentially, it was dried over magnesium sulfateanhydrous. After the drying agent was filtered off, the organic layerwas concentrated under reduced pressure. The obtained residue waspurified by silica gel column chromatography (Kanto Chemical, commercialname Silica gel 60N, granular, neutral, 0.040 mm-0.100 mm;n-heptane:ethyl acetate=10:1→5:1) to obtain the title compound (9.81 g)as a colorless oil.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.01 (s, 3H), 0.02 (s, 3H), 0.86 (s, 9H),0.94 (d, J=6.8 Hz, 3H), 1.22-1.32 (m, 1H), 1.32 (s, 3H), 1.57-1.65 (m,2H), 1.60 (s, 3H), 1.65-1.79 (m, 1H), 2.32 (dd, J=6.0, 15.2 Hz, 1H),2.43 (dd, J=6.0, 15.2 Hz, 1H), 2.42-2.52 (m, 1H), 3.80 (s, 3H), 4.02 (d,J=6.4 Hz, 2H), 4.01-4.08 (m, 1H), 4.26 (dt, J=1.0, 7.0 Hz, 1H), 4.40 (s,2H), 4.96-5.05 (m, 3H), 5.29 (ddd, J=1.0, 1.6, 10.5 Hz, 1H), 5.41 (dt,J=1.6, 17.5 Hz, 1H), 5.62 (brt, J=6.2 Hz, 1H), 5.69 (ddd, J=8.0, 10.4,17.2 Hz, 1H), 5.88 (ddd, J=7.0, 10.5, 17.5 Hz, 1H), 5.90 (s, 1H),6.85-6.89 (m, 2H), 7.23-7.26 (m, 2H), 7.26-7.37 (m, 3H), 7.48-7.52 (m,2H); 100 MHz ¹³C-NMR (CDCl₃) δ (ppm) −4.83, −4.76, 12.65, 16.63, 17.85,22.32, 25.73, 31.21, 32.28, 39.94, 42.72, 55.09, 65.60, 69.21, 71.53,81.37, 82.18, 87.72, 102.16, 113.65, 115.17, 118.64, 126.03, 126.67,128.14, 129.00, 129.27, 130.25, 132.66, 135.33, 137.90, 139.83, 159.08,170.36; IR (neat) 2957, 2931, 2856, 1736, 1612, 1250, 1173, 1090, 1065,1036, 836 cm⁻¹; HRMS C₄₀H₅₈NaO₇Si (M+Na⁺) Calcd: 701.3849, Found:701.3824; [α]_(D) ²³−2.69 (c 1.09, CHCl₃)

(2) Synthesis of(2S,3aS,4E,6S,7S,13aR)-11-{[tert-butyl(dimethyl)silyl]oxy}-7-{(1E)-3-[(4-methoxybenzyl)oxy]-1-methylprop-1-en-1-yl}-6,13a-dimethyl-2-phenyl-3a,6,7,10,11,12,13,13a-octahydro-9H-[1,3]dioxolo[4,5-f]oxacyclododecin-9-one(P17)

Anhydrous toluene distilled by using benzophenone ketyl immediatelybefore use under Ar atmosphere was used in this step.

(1S,2E)-4-[(4-methoxybenzyl)oxy]-2-methyl-1-[(1S)-1-methylprop-2-en-1-yl]but-2-en-1-yl3-{[tert-butyl(dimethyl)silyl]oxy}-5-[(2S,4R,5S)-4-methyl-2-phenyl-5-vinyl-1,3-dioxolan-4-yl]pentanoate(1.10 g, 1.63 mmol) and 2,6-di-tert-butyl-4-methylphenol (35.9 mg, 0.16mmol) were dissolved in anhydrous toluene under Ar atmosphere. Thereaction solution was heated to reflux for one hour. An anhydroustoluene (330 ml) solution of the second generation Hoveyda-Grubbscatalyst;[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(O-isopropoxyphenylmethylene)ruthenium(102 mg, 0.16 mmol) was added to the reaction solution. After thereaction solution was heated to reflux for five hours, it was cooled toroom temperature. The reaction solution was filtered with silica gel(Fuji Silysia, commercial name Chromatorex, NH, 200-350 mesh) and thefiltrate was concentrated under reduced pressure. The obtained residuewas purified by silica gel column chromatography (Kanto Chemical,commercial name Silica gel 60N, granular, neutral, 0.040 mm-0.100 mm;n-heptane:ethyl acetate=8:1) to obtain the title compound (486.1 mg) asa white solid. This was obtained as a colorless needle byrecrystallization (hexane:ethyl acetate).

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.07 (s, 3H), 0.09 (s, 3H), 0.88 (s, 9H),0.91 (d, J=6.8 Hz, 3H), 1.38 (s, 3H), 1.38-1.46 (m, 1H), 1.48-1.56 (m,1H), 1.61 (s, 3H), 1.62-1.68 (m, 1H), 1.99-2.10 (m, 1H), 2.31 (dd,J=10.4, 14.7 Hz, 1H), 2.52-2.58 (m, 1H), 2.58 (dd, J=4.4, 14.7 Hz, 1H),3.81 (s, 3H), 3.92-4.01 (m, 1H), 4.04 (d, J=6.4 Hz, 2H), 4.19 (d, J=9.4Hz, 1H) 4.42 (s, 2H), 4.97 (d, J=10.8 Hz, 1H), 5.41 (dd, J=9.8, 15.2 Hz,1H), 5.63 (dd, J=9.4, 15.2 Hz, 1H), 5.74 (brt, J=6.4 Hz, 1H), 5.91 (s,1H), 6.87-6.90 (m, 2H), 7.24-7.26 (m, 2H), 7.36-7.40 (m, 3H), 7.49-7.51(m, 2H); 100 MHz ¹³C-NMR (CDCl₃) δ (ppm) −4.73, −4.61, 11.50, 16.72,17.80, 22.69, 25.62, 31.44, 34.31, 39.97, 43.98, 55.02, 65.45, 71.51,71.69, 81.55, 83.44, 85.18, 101.07, 113.61, 126.62, 128.18, 128.41,129.12, 129.21, 129.47, 130.05, 134.39, 137.55, 137.63, 159.05, 168.89;IR (neat) 2932, 2856, 1735, 1513, 1460, 1247, 1066, 1034, 1006, 978, 836cm⁻¹; HRMS C₃₈H₅₄NaO₇Si (M+Na⁺) Calcd: 673.3536, Found: 673.3505;[α]_(D) ²⁸−14.8 (c 1.03, CHCl₃)

[Step 9]

Synthesis of (2E)-3-((2S,3aS,4E,6S,7S,13aR)-11-{[tert-butyl(dimethyl)silyl]oxy}-6,13a-dimethyl-9-oxo-2-phenyl-6,7,9,10,11,12,13,13a-octahydro-3aH-[1,3]dioxolo[4,5-f]oxacyclododecin-7-yl)but-2-enal

(1) Synthesis of(2S,3aS,4E,6S,7S,13aR)-11-{[tert-butyl(dimethyl)silyl]oxy}-7-{(1E)-3-hydroxy-1-methylprop-1-en-1-yl}-6,13a-dimethyl-2-phenyl-3a,6,7,10,11,12,13,13a-octahydro-9H-[1,3]dioxolo[4,5-f]oxacyclododecin-9-one(P18)

(2S,3aS,4E,6S,7S,13aR)-11-{[tert-butyl(dimethyl)silyl]oxy}-7-{(1E)-3-[(4-methoxybenzyl)oxy]-1-methylprop-1-en-1-yl}-6,13a-dimethyl-2-phenyl-3a,6,7,10,11,12,13,13a-octahydro-9H-[1,3]dioxolo[4,5-f]oxacyclododecin-9-one(710 mg, 1.09 mmol) was dissolved in dichloromethane (14.0 ml) and aphosphate buffer (pH=7, 1.40 ml).2,3-Dichloro-5,6-dicyano-1,4-benzoquinone (297 mg, 1.31 mmol) was addedto the reaction solution at 0° C. and stirred for three hours.2,3-Dichloro-5,6-dicyano-1,4-benzoquinone (74.3 mg, 0.33 mmol) was addedand the reaction solution was stirred at 0° C. for one and a half hour.After 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (74.3 mg, 0.33 mmol) wasadded and the reaction solution was stirred at 0° C. for further 30minutes, the reaction solution was filtered with silica gel (FujiSilysia, commercial name Chromatorex, NH, 200-350 mesh). The filtratewas washed with a saturated sodium hydrogen carbonate aqueous solution,distilled water and brine sequentially and dried over anhydrousmagnesium sulfate. After the drying agent was filtered off, the organiclayer was concentrated under reduced pressure. The obtained residue waspurified by silica gel column chromatography (Kanto Chemical, commercialname Silica gel 60N, granular, neutral, 0.040 mm-0.100 mm;n-heptane:ethyl acetate=4:1→3:1) to obtain the title compound (460 mg)as a colorless needle.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.08 (s, 3H), 0.09 (s, 3H), 0.88 (s, 9H),0.90 (d, J=6.8 Hz, 3H), 1.25 (t, J=5.6 Hz, 1H), 1.38-1.46 (m, 2H), 1.39(s, 3H), 1.60-1.68 (m, 1H), 1.65 (s, 3H), 2.00-2.09 (m, 1H), 2.32 (dd,J=10.2, 14.6 Hz, 1H), 2.52-2.59 (m, 1H), 2.59 (dd, J=4.2, 14.6 Hz, 1H),3.92-4.01 (m, 1H), 4.16-4.27 (m, 3H), 4.95 (d, J=10.4 Hz, 1H), 5.41 (dd,J=9.8, 15.2 Hz, 1H), 5.64 (dd, J=9.6, 15.2 Hz, 1H), 5.75 (brt, J=5.8 Hz,1H), 5.91 (s, 1H), 7.36-7.41 (m, 3H), 7.49-7.52 (m, 2H); 100 MHz ¹³C-NMR(CDCl₃) δ (ppm) −4.67, −4.56, 11.24, 16.69, 17.88, 22.75, 25.67, 31.50,34.33, 39.92, 43.98, 58.64, 71.49, 81.86, 83.55, 85.25, 101.15, 126.70,128.28, 129.25, 129.56, 130.77, 133.52, 137.58, 169.25; IR (KBr) 3232,2937, 2858, 1730, 1243, 1107, 1082, 1067, 1007, 973, 836, 776, 700 cm-1;HRMS C₃₀H₄₆NaO₆Si (M+Na⁺) Calcd: 553.2961, Found: 553.2948; [α]_(D)²³−11.5 (c 1.02, CHCl₃)

(2) Synthesis of(2S,3aS,4E,6S,7S,13aR)-11-{[tert-butyl(dimethyl)silyl]oxy}-7-[(E)-2-formyl-1-methyleth-1-en-1-yl]-6,13a-dimethyl-2-phenyl-3a,6,7,10,11,12,13,13a-octahydro-9H-[1,3]dioxolo[4,5-f]oxacyclododecin-9-one(P19)

Dess-Martin reagent (565 mg, 1.33 mmol) was added to a dichloromethane(12.0 ml) solution of(2S,3aS,4E,6S,7S,13aR)-11-{[tert-butyl(dimethyl)silyl]oxy}-7-{(1E)-3-hydroxy-1-methylprop-1-en-1-yl}-6,13a-dimethyl-2-phenyl-3a,6,7,10,11,12,13,13a-octahydro-9H-[1,3]dioxolo[4,5-f]oxacyclododecin-9-one(587 mg, 1.11 mmol). The reaction solution was stirred at roomtemperature for 30 minutes. The reaction solution was diluted with etherand washed with a saturated sodium hydrogen carbonate aqueous solutioncontaining sodium sulfite, water and brine sequentially and dried overanhydrous magnesium sulfate. After the drying agent was filtered off,the organic layer was concentrated under reduced pressure. The obtainedresidue was purified by silica gel column chromatography (KantoChemical, commercial name Silica gel 60N, granular, neutral, 0.040mm-0.100 mm; n-heptane:ethyl acetate=10:1→8:1→6:1) to obtain the titlecompound (587 mg) as a white solid. This was obtained as a colorlesscrystal by recrystallization (hexane:ethyl acetate).

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.09 (s., 3H), 0.10 (s, 3H), 0.89 (s,9H), 0.95 (d, J=6.8 Hz, 3H), 1.38-1.50 (m, 2H), 1.40 (s, 3H), 1.63-1.70(m, 1H), 1.96-2.10 (m, 1H), 2.17 (d, J=1.2 Hz, 3H), 2.37 (dd, J=9.6,14.4 Hz, 1H), 2.60 (dd, J=4.0, 14.4 Hz, 1H), 2.61 (m, 1H), 3.93-4.03 (m,1H), 4.20 (d, J=9.6 Hz, 1H), 4.92 (d, J=10.4 Hz, 1H), 5.41 (dd, J=9.2,15.2 Hz, 1H), 5.69 (dd, J=9.6, 15.2 Hz, 1H), 5.92 (s, 1H), 6.07 (dd,J=1.2, 7.6 Hz, 1H), 7.34-7.42 (m, 3H), 7.48-7.52 (m, 2H), 10.0 (d, J=7.6Hz, 1H); 100 MHz ¹³C-NMR (CDCl₃) δ (ppm) −4.70, −4.60, 13.20, 16.31,17.87, 22.66, 25.64, 31.46, 34.14, 40.40, 43.19, 71.13, 80.52, 83.52,85.00, 101.22, 126.63, 128.28, 129.25, 129.84, 130.58, 136.19, 137.59,155.92, 168.80, 190.56; IR (KBr) 2954, 2935, 2880, 2858, 2787, 2755,1739, 1674, 1461, 1401, 1240, 1224, 1101, 1004, 980, 830, 775, 698 cm⁻¹;HRMS C₃₀H₄₄NaO₆Si (M+Na⁺) Calcd: 551.2805, Found: 551.2803; [α]_(D)²⁷+4.00 (c 1.06, CHCl₃)

[Step 10]

Synthesis of (3S)-4-(benzyloxy)-3-methylbutan-1-ol

(1) Synthesis of methyl (2R)-3-(benzyloxy)-2-methylpropanoate (P20)

This reaction was performed with reference to the literature (Widmer,U.; Synthesis, 1987, 568-570.).

Methyl(R)-3-hydroxy-isobutyrate (6.30 g, 53.3 mmol) was dissolved in amixed solution (180 ml) of dichloromethane-cyclohexane (1:1), followedby addition of benzyl-2,2,2-trichloroacetimidate (12.0 ml, 64.6 mmol) atroom temperature. After trifluoromethanesulfonic acid (0.80 ml, 5.40mmol) was added dropwise to this, the reaction solution was stirred atroom temperature for three hours. After the reaction solution wasdiluted with dichloromethane, it was washed with a sodium hydrogencarbonate aqueous solution and brine. After the organic layer was driedover anhydrous sodium sulfate and the drying agent was filtered off, theorganic layer was concentrated under reduced pressure. The obtainedresidue was purified by silica gel column chromatography (Merck,commercial name Silica Gel 60, 230-400 mesh; hexane:ethyl acetate=30:1)and the title compound (9.79 g) was obtained as a colorless oil.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 1.19 (d, J=7.2 Hz, 3H), 2.74-2.84 (m,1H), 3.50 (dd, J=6.0, 9.2 Hz, 1H), 3.66 (dd, J=7.2, 9.2 Hz, 1H), 3.69(s, 3H), 4.52 (s, 2H), 7.20-7.37 (m, 5H); 100 MHz ¹³C-NMR (CDCl₃) δ(ppm) 13.91, 40.12, 51.65, 71.89, 73.02, 127.51, 127.53, 128.28, 138.09,175.24; IR (neat) 2977, 2946, 2862, 2362, 2323, 1738, 1458, 1202 cm⁻¹;HRMS C₁₂H₁₆NaO₃ (M+Na⁺) Calcd: 231.0997, Found: 231.0989; [α]_(D)²⁸−10.9 (c 2.10, CHCl₃)

(2) Synthesis of({[(2S)-4-methoxy-2-methylbut-3-en-1-yl]oxy}methyl)benzene (P21)

This reaction was performed with reference to the literature (Rich, D.H.; Sun, E. T.; Boparai, A. S.; J. Org. Chem., 1978, 43(18) 3624-3626.Feng, X.; Edstrom, E. D., Tetrahedron Asymmetry, 1999, 10(1), 99-105.Gibson, S. E.; Guillo, N.; Middleton, R. J.; Thuilliez, A.; Tozer, M.J.; J. Chem. Soc. Perkin Trans., 1997, 1, 447-455.).

1.0M diisobutylaluminum hydride toluene solution (3.78 ml) was addeddropwise to a toluene (32 ml) solution of methyl(2R)-3-(benzyloxy)-2-methylpropanoate (0.81 g, 3.88 mmol) at −78° C. andthe reaction solution was stirred at the same temperature for 1.5 hours.After addition of methanol (0.5 ml, 12.3 mmol), the reaction solutionwas warmed to room temperature. The reaction solution was stirred forfurther two hours. The reaction solution was filtered through celite andthe filtrate was dried over sodium sulfate. The drying agent wasfiltered off, the solvent was evaporated under reduced pressure andaldehyde was obtained as a crude product.

After potassium tert-butoxide (0.76 g, 6.76 mmol) was added to a THF (20ml) solution of (methoxymethyl)triphenylphosphonium chloride (2.54 g,7.40 mmol) while ice cooling, the mixture was stirred at roomtemperature for 30 minutes. Subsequently a THF (5 ml) solution of theabove aldehyde was added dropwise to this reaction solution at roomtemperature and stirred for 12 hours. After the reaction solution wasdiluted with ethyl acetate, it was washed with water and brine. Afterthe organic layer was dried over anhydrous magnesium sulfate and thedrying agent was filtered off, the organic layer was concentrated underreduced pressure. The obtained residue was purified by silica gel columnchromatography (Merck, commercial name Silica Gel 60, 230-400 mesh;hexane:ethyl acetate=100:1) to obtain the title compound (0.44 g) as acolorless oil. Title compound was determined to be a mixture of E:Z=2:1by ¹H-NMR.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 1.01 (d, J=6.8 Hz, 0.9H), 1.04 (d, J=6.8Hz, 2.1H), 2.36-2.47 (m, 0.7H), 2.92-3.00 (m, 0.3H), 3.22-3.37 (m, 2H),3.50 (s, 2.1H), 3.58 (s, 0.9H), 4.23 (dd, J=6.4, 9.2 Hz, 0.3H), 4.52 (s,2.0H), 4.65 (dd, J=8.0, 12.8 Hz, 0.7H), 5.89 (dd, J=0.8, 6.4 Hz, 0.3H),6.35 (dd, J=0.8, 12.8 Hz, 0.7H), 7.25-7.36 (m, 5H); 100 MHz ¹³C-NMR(CDCl₃) δ (ppm) 17.97, 18.43, 29.60, 33.22, 55.67, 59.50, 72.67, 72.88,75.24, 76.16, 105.60, 109.51, 127.34, 127.45, 127.53, 128.25, 128.31,138.72, 138.92, 146.35, 147.31; IR (neat) 3060, 3033, 2956, 2935, 2856,1655, 1454, 1207, 1098, 737 cm⁻¹; HRMS C₁₃H₁₈NaO₂ (M+Na⁺) Calcd:229.1204, Found: 229.1207.

(3) Synthesis of (3S)-4-(benzyloxy)-3-methylbutan-1-ol (P22)

This reaction was performed with reference to the literature (Gibson, S.E.; Guillo, N.; Middleton, R. J.; Thuilliez, A.; Tozer, M. J.;. J. Chem.Soc., Perkin Trans., 1997, 1, 447-455.).

Water (10 ml) and formic acid (1 ml) were added to({[(2S)-4-methoxy-2-methylbut-3-en-1-yl]oxy}methyl)benzene (3.73 g, 18mmol) and the reaction solution was stirred at room temperature for 10minutes. The reaction solution was diluted with ethyl acetate and washedwith water, a sodium hydrogen carbonate aqueous solution and brinesequentially. After the organic layer was dried over anhydrous sodiumsulfate and the drying agent was filtered off, the organic layer wasconcentrated under reduced pressure.

Sodium borohydride (0.68 g, 18 mmol) was added to a methanol (40 ml)solution of the obtained crude product while ice cooling. After thereaction solution was stirred at the same temperature for 10 minutes,acetone was added thereto. The reaction solution was diluted with ethylacetate and washed with brine. After the organic layer was dried overanhydrous sodium sulfate and the drying agent was filtered off, theorganic layer was concentrated under reduced pressure. The obtainedresidue was purified by silica gel column chromatography (Merck,commercial name Silica Gel 60, 230-400 mesh; hexane:ethyl acetate=5:1)to obtain the title compound (3.30 g) as a colorless oil.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.95 (d, J=7.2 Hz, 3H), 1.51-1.68 (m,2H), 1.89-1.99 (m, 1H), 3.31 (dd, J=7.6, 9.2 Hz, 1H), 3.39 (dd, J=4.8,9.2 Hz, 1H), 3.61-3.75 (m, 2H), 4.52 (s, 2H), 7.26-7.37 (m, 5H); 100 MHz¹³C-NMR (CDCl₃) δ (ppm) 17.51, 31.18, 37.74, 60.88, 73.08, 75.96,127.57, 127.58, 128.31, 137.99; IR (neat) 3389, 3086, 3058, 3030, 2951,2929, 2872, 1454, 1363, 1205, 1095, 738, 698 cm⁻¹; HRMS C₁₂H₁₈NaO₂(M+Na⁺) Calcd: 217.1204, Found: 217.1190; [α]_(D) ²⁹−4.52 (c 2.41,CHCl₃)

[Step 11]

Synthesis of5-{[(3S)-4-(benzyloxy)-3-methylbutyl]sulfonyl}-1-phenyl-1H-tetrazole

(1) Synthesis of5-{[(3S)-4-(benzyloxy)-3-methylbutyl]thio}-1-phenyl-1H-tetrazole (P23)

This reaction was performed with reference to the literature (Mitsunobu,O., Synthesis, 1981, 1-28.).

To a THF (60 ml) solution of (3S)-4-(benzyloxy)-3-methylbutan-1-ol (2.76g, 14.2 mmol), 5-mercapto-1-phenyltetrazole (3.03 g, 17 mmol),triphenylphosphine (4.47 g, 17 mmol) and diethyl azodicarboxylate 40%toluene solution (8.04 ml, 18 mmol) were added while ice cooling. Thereaction solution was warmed to room temperature and was stirred forthree hours. After the reaction solution was diluted with ethyl acetate,it was washed with distilled water and brine. After the organic layerwas dried over anhydrous sodium sulfate and the drying agent wasfiltered off, the organic layer was concentrated under reduced pressure.The obtained residue was purified by silica gel column chromatography(Merck, commercial name Silica Gel 60, 230-400 mesh; hexane:ethylacetate=15:1→5:1) to obtain the title compound (5.38 g) as a colorlessoil.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.99 (d, J=6.8 Hz, 3H), 1.65-1.74 (m,1H), 1.92-2.03 (m, 2H), 3.26-3.52 (m, 4H), 4.49 (s, 2H), 7.21-7.35 (m,5H), 7.49-7.58 (m, 5H); 100 MHz ¹³C-NMR (CDCl₃) δ (ppm) 16.72, 31.23,32.82, 33.13, 73.02, 75.07, 123.78, 127.48, 128.30, 129.70, 129.99,133.67, 138.39, 154.37; IR (neat) 3067, 3036, 2956, 2925, 2858, 1598,1499, 1386, 1092, 761, 696 cm⁻¹; HRMS C₁₉H₂₃N₄OS (M+H⁺) Calcd: 355.1593,Found: 355.1583; [α]_(D) ²⁷−2.62 (c 1.56, CHCl₃)

(2) Synthesis of5-{[(3S)-4-(benzyloxy)-3-methylbutyl]sulfonyl}-1-phenyl-1H-tetrazole(P24)

This reaction was performed with reference to the literature (Shultz, H.S.; Freyermuth, H. B.; Buc, S. R., J. Org. Chem., 1963, 28(4),1140-1142.).

About 30% hydrogen peroxide solution (16.5 ml, 146 mmol) of hexaammoniumheptamolybdate tetrahydrate (1.80 g, 1.46 mmol) was added to an ethanol(20 ml) solution of5-{[(3S)-4-(benzyloxy)-3-methylbutyl]thio}-1-phenyl-1H-tetrazole (5.17g, 14.6 mmol) at room temperature. After the reaction solution wasstirred at room temperature for 24 hours, it was diluted with ethylacetate and washed with water and brine. After the organic layer wasdried over anhydrous sodium sulfate and the drying agent was filteredoff, the organic layer was concentrated under reduced pressure. Theobtained residue was purified by silica gel column chromatography(Merck, commercial name Silica Gel 60, 230-400 mesh; hexane:ethylacetate=10:1) to obtain the title compound (5.68 g) as a colorless oil.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.98 (d, J=6.8 Hz, 3H), 1.83-2.13 (m,3H), 3.29 (dd, J=6.8, 9.6 Hz, 1H), 3.40 (dd, J=4.8, 9.6 Hz, 1H),3.74-3.87 (m, 2H), 4.49 (s, 2H), 7.25-7.36 (m, 5H), 7.55-7.68 (m, 5H);100 MHz ¹³C-NMR (CDCl₃) δ (ppm) 16.72, 26.23, 32.49, 54.30, 73.11,74.70, 125.05, 127.55, 127.62, 128.39, 129.63, 131.37, 132.99, 138.09,153.38; HRMS C₁₉H₂₃N₄O₃S (M+H⁺) Calcd: 387.1491, Found: 387.1468;[α]_(D) ²⁶−5.28 (c 1.00, CHCl₃)

[Step 12]

Synthesis of(4R)-4-benzyl-3-[(2R,3S)-3-hydroxy-2-methylpentanoyl]-1,3-oxazolidin-2-one

(1) Synthesis of (4R)-4-benzyl-3-propionyl-1,3-oxazolidin-2-one (P26)

This reaction was performed with reference to the literature (Gage, J.R.; Evans, D. A., Organic Synthesis, 1989, 68, 83-91. Chan, P. C.-M.;Chong. J. M.; Kousha, K., Tetrahedron, 1994, 150(9), 2703-2714.).

1.57M n-butyllithium hexane solution (96 ml, 150 mmol) was slowly addeddropwise to a THF (400 ml) solution of (R)-4-benzyl-2-oxazolidinone(25.36 g, 143 mmol) at −78° C. under stirring. Subsequently propionylchloride (13.7 ml, 157 mmol) was added at once and the reaction solutionwas stirred at the same temperature for 30 minutes. After the reactionsolution was warmed to room temperature for 30 minutes, it was dilutedwith ethyl acetate and washed with water and brine. The organic layerwas dried over anhydrous sodium sulfate. After the drying agent wasfiltered off, the organic layer was concentrated under reduced pressure.The obtained residue was purified by silica gel column chromatography(Merck, commercial name Silica Gel 60, 230-400 mesh; heptane:ethylacetate=5:1) to obtain the title compound (36.58 g) as a white solid.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 1.21 (t, J=7.2 Hz, 3H), 2.77 (dd, J=9.6,13.2 Hz, 1H), 2.88-3.05 (m, 2H), 3.31 (dd, J=3.2, 13.2 Hz, 1H),4.11-4.23 (m, 2H), 4.65-4.70 (m, 1H), 7.20-7.36 (m, 5H); 100 MHz ¹³C-NMR(CDCl₃) δ (ppm) 8.52, 29.42, 38.15, 55.39, 66.44, 127.56, 129.17,129.64, 135.55, 164.52, 174.30; IR (KBr) 3082, 3026, 2984, 2942, 2872,2360, 2338, 1787, 1702, 1496 cm⁻¹; HRMS C₁₃H₁₅NNaO₃ (M+Na⁺) Calcd:256.0950, Found: 256.0928; [α]_(D) ²⁷−101.20 (c 1.11, CH₃CH₂OH), [α]_(D)²⁵−63.9 (c 1.00, CHCl₃)

(2) Synthesis of(4R)-4-benzyl-3-[(2R,3S)-3-hydroxy-2-methylpentanoyl]-1,3-oxazolidin-2-one(P27)

This reaction was performed with reference to the literature (Gage, J.R.; Evans, D. A., Organic Synthesis, 1989, 68, 83-91. Chan, P. C.-M.;Chong. J. M.; Kousha, K., Tetrahedron, 1994, 50(9), 2703-2714. Cane, D.E.; Tan, W.; Ott, W. R., J. Am. Chem. Soc., 1993, 115(2) 527-535.).

To a dichloromethane (120 ml) solution of(4R)-4-benzyl-3-propionyl-1,3-oxazolidin-2-one (17.22 g, 73.8 mmol), 1Mdibutylboron triflate dichloromethane solution (150 ml, 150 mmol) andtriethylamine (24.4 ml, 179 mmol) were added dropwise while ice cooling.Subsequently the reaction solution was cooled to −78° C. andpropioaldehyde (10.5 ml, 145 mmol) was added dropwise over five minutes.After the reaction solution was stirred at the same temperature for onehour, it was stirred at 0° C. for two hours. The reaction solution wasice cooled again, followed by addition of a mixed solution of phosphatebuffer (pH=7)-distilled water (1:3, 160 ml). After further cooled to−10° C., a mixed solution of methanol −30% hydrogen peroxide solution(2:1, 120 ml) was added thereto, and the mixture was stirred for onehour. After the reaction solution was concentrated under reducedpressure, it was diluted with diethyl ether and washed with a saturatedsodium hydrogen carbonate aqueous solution and brine. The organic layerwas dried over anhydrous sodium sulfate. After the drying agent wasfiltered off, the organic layer was concentrated under reduced pressure.The obtained residue was purified by silica gel column chromatography(Merck, commercial name Silica Gel 60, 230-400 mesh; hexane:ethylacetate=5:1) to obtain the title compound (16.4 g) as a white solid.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.98 (t, J=7.3 Hz, 3H), 1.25 (d, J=6.9Hz, 3H), 1.40-1.64 (m, 2H), 2.77 (dd, J=9.5, 13.4 Hz, 1H), 2.88 (brs,1H), 3.25 (dd, 3.1, 13.4 Hz, 1H), 3.80 (dq, J=2.7, 6.8 Hz, 1H),3.85-3.89 (m, 1H), 4.17-4.25 (m, 2H), 4.68-4.74 (m, 1H), 7.19-7.36 (m,5H); 100 MHz ¹³C-NMR (CDCl₃) δ (ppm) 10.24, 10.41, 26.72, 37.75, 41.64,55.07, 66.13, 72.93, 127.39, 128.93, 129.38, 134.99, 152.98, 177.53; IR(KBr) 3654, 3526, 3375, 3086, 3030, 2976, 2937, 2341, 1760, 1700, 1455,1376, 1265, 1220, 1117, 1071, 971, 930, 851, 767 cm⁻¹; HRMS C₁₆H₂₁NNaO₄(M+Na⁺) Calcd: 314.1368, Found: 314.1381; [α]_(D) ²⁸−52.2 (c 1.00,CHCl₃)

[Step 13]

Synthesis of (2R,3S)-2-methyl-3-[(trietylsilyl)oxy]pentanal

(1) Synthesis of(2R,3S)—N-methoxy-N,2-dimethyl-3-[(triethylsilyl)oxy]pentanamide (P28)

This reaction was performed with reference to the literature (Nahm, S.;Weinreb, S. M., Tetrahedron Lett., 1981, 22(39), 3815-3818. Cane, D. E.;Tan, W.; Ott, W. R., J. Am. Chem. Soc., 1993, 115(2) 527-535.DiBattista, J. P.; Webster, F. X., Bioorg. Med. Chem., 1996, 4(3),423-428.).

2M trimethylaluminum toluene solution (90 ml, 180 mmol) was added to aTHF (180 ml) solution of N,O-dimethylhydroxyamine hydrochloride (18.3 g,187.6 mmol) at −10° C. under stirring. After the reaction solution waswarmed to 0° C. and stirred for 10 minutes, it was warmed to roomtemperature and further stirred for 30 minutes. The reaction solutionwas cooled to −10° C. again, and a THF-dichloromethane solution (4:5,180 ml) of(4R)-4-benzyl-3-[(2R,3S)-3-hydroxy-2-methylpentanoyl]-1,3-oxazolidin-2-one(21.85 g, 75 mmol) was added dropwise to the reaction solution. Afterthe reaction solution was warmed to 0° C. and stirred for two hours, amixed solution of dichloromethane −0.5N hydrochloric acid (1:1, 80 ml)was added slowly, and the mixture was stirred for one hour at 0° C. Thereaction solution was filtered through celite and the filtrate was driedover anhydrous sodium sulfate. After the drying agent was filtered off,the organic layer was concentrated under reduced pressure. The obtainedresidue was purified by silica gel column chromatography (Merck,commercial name Silica Gel 60; heptane:ethyl acetate=1.5:1) to obtain(2R,3S)-3-hydroxy-N-methoxy-N,2-dimethylpentanamide (13.0 g) as acolorless oil.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.96 (t, J=7.2 Hz, 3H), 1.16 (d, J=7.2Hz, 3H), 1.34-1.45 (m, 1H), 1.53-2.04 (m, 1H), 2.84-2.94 (m, 1H), 3.19(s, 3H), 3.71 (s, 3H), 3.74-3.80 (m, 1H); 100 MHz ¹³C-NMR (CDCl₃) δ(ppm) 10.08, 10.21, 26.64, 31.73, 38.09, 61.35, 72.95, 177.21; IR (neat)3433, 2971, 2943, 2881, 2361, 2337, 1638, 1461, 993 cm⁻¹; HRMSC₈H₁₇NNaO₃ (M+Na⁺) Calcd: 198.1106, Found: 198.1100; [α]_(D) ²⁵−17.3 (c1.11, CHCl₃)

The above (2R,3S)-3-hydroxy-N-methoxy-N,2-dimethylpentanamide (7.3 g,41.7 mmol) was dissolved in dichloromethane (150 ml), followed byaddition of 2,6-lutidine (10.2 ml, 87 mmol) and trifluoromethanesulfonicacid triethylsilyl ester (14.1 ml, 62 mmol) while ice cooling, and thereaction solution was stirred for five hours. The reaction solution wasdiluted with dichloromethane and washed with a saturated ammoniumchloride aqueous solution and brine. After the organic layer was driedover anhydrous sodium sulfate and the drying agent was filtered off, theorganic layer was concentrated under reduced pressure. The obtainedresidue was purified by silica gel column chromatography (Merck,commercial name Silica Gel 60, 230-400 mesh; hexane:ethyl acetate=10:1)to obtain the title compound (11.95 g) as a colorless oil.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.62 (q, J=8.0 Hz, 6H), 0.90 (t, J=7.2Hz, 3H), 0.98 (t, J=8.0 Hz, 9H), 1.17 (d, J=6.8 Hz, 3H), 1.40-1.59 (m,2H), 2.90-3.08 (m, 1H), 3.18 (s, 3H), 3.69 (s, 3H), 3.90 (dt, J=4.8, 8.4Hz, 1H); 100 MHz ¹³C-NMR (CDCl₃) δ (ppm) 4.96, 6.74, 8.60, 14.35, 28.25,31.86, 40.21, 61.16, 74.50, 176.51; IR (neat) 3483, 2958, 2918, 2879,1743, 1663, 1460, 1384, 1118, 1049, 1007, 857, 741 cm⁻¹; HRMSC₁₄H₃₂NO₃Si (M+H⁺) Calcd: 290.2151, Found: 290.2150; [α]_(D) ²⁷−7.39 (c1.04, CHCl₃)

(2) Synthesis of (2R,3S)-2-methyl-3-[(triethylsilyl)oxy]pentanal (P29)

This reaction was performed with reference to the literature (Nahm, S.;Weinreb, S. M., Tetrahedron Lett., 1981, 22(39), 3815-3818).

To a THF (20 ml) solution of(2R,3S)—N-methoxy-N,2-dimethyl-3-[(triethylsilyl)oxy]pentanamide (1.06g, 3.66 mmol), 1M diisobutylaluminum hydride toluene solution (18 ml)was added dropwise at −78° C. and it was stirred at the same temperaturefor one hour. After 5% hydrogen chloride methanol solution (2 ml) wasadded to the reaction solution, the mixture was warmed to roomtemperature and stirred for one hour. The reaction solution was filteredthrough celite and the filtrate was concentrated under reduced pressure.The obtained residue was purified by silica gel column chromatography(Merck, commercial name Silica Gel 60, 230-400 mesh; hexane:ethylacetate=40:1) to obtain the title compound (0.75 g) as a colorless oil.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.60 (q, J=8.0 Hz, 6H), 0.89 (t, J=7.2Hz, 3H), 0.94 (t, J=8.0 Hz, 9H), 1.06 (d, J=6.8 Hz, 3H), 1.43-1.62 (m,2H), 2.40-2.49 (m, 1H), 4.05 (ddd, J=3.4, 6.4, 9.6 Hz, 1H), 9.77 (s,1H); 100 MHz ¹³C-NMR (CDCl₃) δ (ppm) 5.01, 6.74, 7.43, 10.01, 27.47,50.82, 73.33, 205.26; IR (neat) 3455, 2958, 2914, 2872, 2715, 2356,2341, 1726, 1460, 1239, 1011, 740 cm⁻¹; HRMS C₁₂H₂₆NaO₂Si (M+Na⁺) Calcd:253.1600, Found: 253.1596; [α]_(D) ²⁴−55.3 (c 1.19, CHCl₃)

[Step 14]

Synthesis of(2S,4E,6S,7S)-2,6-dimethyl-7-[(triethylsilyl)oxy]non-4-en-1-ol

(1) Synthesis of{[(1S,2S,3E,6S)-7-(benzyloxy)-1-ethyl-2,6-dimethylhept-3-en-1-yl]oxy}(triethyl)silane(P30)

This reaction was performed with reference to the literature (Blakemore,P. R.; Cole, W. J.; Kociensky, P. J.; Morley, A., Synlett, 1998,26-28.).

THF distilled by using lithium aluminum hydride immediately before usewas used at this step.

15% toluene solution (0.65 ml) of potassium bis(trimethylsilyl)amide wasadded dropwise to THF (2 ml) solution of5{[(3S)-4-(benzyloxy)-3-methylbutyl]sulfonyl}-1-phenyl-1H-tetrazole (100mg, 0.244 mmol) at −78° C. and it was stirred at the same temperaturefor one hour. Subsequently a THF (0.5 ml) solution of(2R,3S)-2-methyl-3-[(triethylsilyl)oxy]pentanal (112 mg, 0.49 mmol) wasadded dropwise to the reaction solution at −78° C. and it was stirredfor one hour. After the reaction solution was warmed to roomtemperature, an appropriate amount of water was added. The reactionsolution was diluted with ethyl acetate and washed with brine. After theorganic layer after was dried over anhydrous sodium sulfate and thedrying agent was filtered off, the organic layer was concentrated underreduced pressure. The obtained residue was purified by silica gel columnchromatography (Merck, commercial name Silica Gel 60, 230-400 mesh;hexane:diethyl ether=100:1) to obtain the title compound (66 mg) as acolorless oil.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.60 (q, J=8.0 Hz, 6H), 0.86 (t, J=7.2Hz, 3H), 0.91 (d, J=6.4 Hz, 3H), 0.94 (d, J=6.8 Hz, 3H), 0.96 (t, J=8.0Hz, 9H), 1.34-1.49 (m, 2H), 1.78-1.93 (m, 2H), 2.11-2.19 (m, 1H),2.20-2.27 (m, 1H), 3.24 (dd, J=6.0, 8.8 Hz, 1H), 3.33 (dd, J=6.0, 8.8Hz, 1H), 3.42 (dt, J=5.2, 5.6 Hz, 1H), 4.49 (s, 2H), 5.31-5.41 (m, 2H),7.26-7.36 (m, 5H); IR (neat) 3086, 3067, 3033, 2958, 2903, 2879, 1500,1456, 1102, 1011, 731 cm⁻¹; HRMS C₂₄H₄₂AgO₂Si (M+Ag⁺) Calcd: 497.2005,Found: 497.1981

100 MHz ¹³C-NMR (CDCl₃) δ (ppm) 5.18, 6.97, 9.35, 16.15, 16.84, 26.81,33.80, 36.83, 41.73, 72.92, 75.33, 77.44, 127.34, 127.44, 127.48,128.23, 134.81, 138.77; [α]_(D) ²⁵−16.07 (c 1.28, CHCl₃′).

(2) Synthesis of(2S,4E,6S,7S)-2,6-dimethyl-7-[(triethylsilyl)oxy]non-4-en-1-ol (P31)

This reaction was performed with reference to the literature (Shimshock,S. J.; Waltermire, R. E.; DeShong, P., J. Am. Chem. Soc., 1991, 113,8791-8796.).

Lithium (39.4 mg, 5.62 mmol) was added to a THF (25 ml) solution ofdi-tert-butyldiphenyl (1.87 g, 7.02 mmol) while ice cooling and thereaction solution was stirred at room temperature for three hours. Thereaction solution turned dark green during stirring. This solution wasadded dropwise to THF (5 ml) solution of{[(1S,2S,3E,6S)-7-benzyloxy-1-ethyl-2,6-dimethylhept-3-en-1-yl]oxy}(triethyl)silane(500 mg, 1.28 mmol) slowly (at such a rate that green of the reactionsolution was maintained) cooled to −78° C. under stirring. After thereaction solution was stirred at the same temperature for three hours, asaturated ammonium chloride aqueous solution was added thereto and themixture was warmed to room temperature. After the reaction solution wasdiluted with ethyl acetate, it was washed with brine. After the organiclayer was dried over anhydrous sodium sulfate and the drying agent wasfiltered off, the organic layer was concentrated under reduced pressure.The obtained residue was purified by silica gel column chromatography(Merck, commercial name Silica Gel 60, 230-400 mesh; hexane:ethylacetate=15:1) to obtain the title compound (296 mg) as a colorless oil.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.60 (q, J=8.0 Hz, 6H), 0.87 (t, J=7.6Hz, 3H), 0.90 (d, J=6.8 Hz, 3H), 0.95 (d, J=7.6 Hz, 3H), 0.96 (t, J=8.0Hz, 9H), 1.34-1.50 (m, 2H), 1.65-1.76 (m, 1H), 1.87-1.94 (m, 1H), 2.09(ddd, 5.6, 5.6, 14.0 Hz, 1H), 2.21-2.29 (m, 1H), 3.42-3.48 (m, 2H), 3.51(dd, J=6.0, 10.4 Hz, 1H), 5.30-5.46 (m, 2H); IR (neat) 3342, 2958, 2916,2877, 1459, 1415, 1380, 1013, 741 cm⁻¹; HRMS C₁₇H₃₆AgO₂Si (M+Ag⁺) Calcd:407.1536, Found: 407.1512

100 MHz ¹³C-NMR (CDCl₃) δ (ppm) 5.11, 6.90, 9.39, 16.05, 16.37, 26.67,35.96, 36.69, 41.71, 67.81, 77.46, 127.57, 134.84; [α]_(D) ²⁵−24.74 (c1.36, CHCl₃).

[Step 15]

Synthesis of(3S,4S,5E,8S)-4,8-dimethyl-9-[(1-phenyl-1H-tetrazol-5-yl)sulfonyl]non-5-en-3-ol

(1) Synthesis of5-{(2S,4E,6S,7S)-2,6-dimethyl-7-[(triethylsilyl)oxy]non-4-en-1-yl}thio)-1-phenyl-1H-tetrazole(P32)

This reaction was performed with reference to the literature (Mitsunobu,O., Synthesis, 1981, 1-28.).

After 5-mercapto-1-phenyltetrazole (237 mg, 1.33 mmol) andtriphenylphosphine (350 mg, 1.33 mmol) were added to a THF (10 ml)solution of(2S,4E,6S,7S)-2,6-dimethyl-7-[(triethylsilyl)oxy]non-4-en-1-ol (334 mg,1.11 mmol) at room temperature, 40% toluene solution of diethylazodicarboxylate (0.63 ml, 1.44 mmol) was added dropwise to while icecooling. After the reaction solution was stirred at room temperature forthree hours, it was diluted with ethyl acetate and washed with distilledwater and brine. After the organic layer was dried over anhydrous sodiumsulfate and the drying agent was filtered off, the organic layer wasconcentrated under reduced pressure. The obtained residue was purifiedby silica gel column chromatography (Merck, commercial name Silica Gel60, 230-400 mesh; hexane:ethyl acetate=40:1) to obtain the titlecompound (457 mg) as a colorless oil.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.59 (q, J=8.0 Hz, 6H), 0.85 (t, J=7.6Hz, 3H), 0.95 (d, J=6.8 Hz, 3H), 0.95 (t, J=8.0 Hz, 9H), 1.03 (d, J=6.0Hz, 3H), 1.32-1.52 (m, 2H), 1.97-2.06 (m, 2H), 2.18-2.29 (m, 2H), 3.28(dd, J=6.4, 12.8 Hz, 1H), 3.41-3.45 (m, 2H), 5.32-5.39 (m, 1H), 5.45(dd, J=7.2, 15.2 Hz, 1H), 7.52-7.60 (m, 5H); IR (neat) 2958, 2883, 1594,1500, 1459, 1383, 1240, 1013, 742 cm⁻¹; HRMS C₂₄H₄₁N₄OSSi (M+H⁺) Calcd:461.2770, Found: 461.2787

100 MHz ¹³C-NMR (CDCl₃) δ (ppm) 5.09, 6.90, 9.40, 16.05, 15.86, 18.80,26.64, 33.09, 38.84, 39.71, 41.68, 77.29, 123.75, 126.09, 129.66,129.95, 133.68, 135.96, 154.56; [α]_(D) ²⁵−22.0 (c 1.51, CHCl₃).

(2) Synthesis of(3S,4S,5E,8S)-4,8-dimethyl-9-[(1-phenyl-1H-tetrazol-5-yl)sulfonyl]non-5-en-3-ol(P33)

This reaction was performed with reference to the literature (Shultz, H.S.; Freyermuth, H. B.; Buc, S. R., J. Org. Chem., 1963, 28(4),1140-1142.).

About 30% hydrogen peroxide solution (1.29 ml, 11.4 mmol) solution ofhexaammonium heptamolybdate tetrahydrate (140 mg, 0.12 mmol) was addedto an ethanol (10 ml) solution of5-{(2S,4E,6S,7S)-2,6-dimethyl-7-[(triethylsilyl)oxy]non-4-en-1-yl}thio)-1-phenyl-1H-tetrazole(444 mg, 1.14 mmol) at room temperature and the reaction solution wasstirred for 24 hours. The reaction solution was diluted with ethylacetate and washed with water and brine. After the organic layer wasdried over anhydrous sodium sulfate and the drying agent was filteredoff, the organic layer was concentrated under reduced pressure. Theobtained residue was purified by silica gel column chromatography(Merck, commercial name Silica Gel 60, 230-400 mesh; hexane:ethylacetate=5:1-1:1) to obtain the title compound (300 mg) as a colorlessoil.

(The title compound crystallized from hexane-ethyl acetate.)

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.95 (t, J=7.3 Hz, 3H), 1.01 (d, J=6.8Hz, 3H), 1.16 (d, J=6.6 Hz, 3H), 1.31-1.42 (m, 1H), 1.42-1.49 (m, 1H),1.49-1.59 (m, 1H), 2.20 (dd, J=6.2, 6.4 Hz, 2H), 2.23-2.31 (m, 1H),2.37-2.47 (m, 1H), 3.34-3.42 (m, 1H), 3.53 (dd, J=7.4, 14.5 Hz, 1H),3.88 (dd, J=4.8, 14.5 Hz, 1H), 5.39-5.47 (m, 1H), 5.50 (dd, J=7.2, 15.6Hz, 1H), 7.57-7.70 (m, 5H); 100 MHz ¹³C-NMR (CDCl₃) δ (ppm) 10.24,14.79, 19.60, 26.92, 28.25, 39.35, 42.14, 60.73, 76.35, 125.02, 126.09,129.55, 131.34, 132.92, 137.02, 153.88; IR (KBr) 3384, 3351, 3316, 2961,2939, 2879, 2356, 2338, 1593, 1499, 1459, 1332, 1156, 1098, 1019, 967,840, 768, 691, 633 cm⁻¹; HRMS C₁₈H₂₇N₄O₃S (M+H⁺) Calcd: 379.1804, Found:379.1806; [α]_(D) ²³−29.5 (c 1.20, CHCl₃)

[Step 16]

Synthesis of5-({(2S)-3-[(2R,3R)-3-((1S,2S)-2-{[diethyl(isopropyl)silyl]oxy}-1-methylbutyl)oxiran-2-yl]-2-methylpropyl}sulfonyl)-1-phenyl-1H-tetrazole)

(1) Synthesis of(2R,3S)-2-((2R,3R)-3-{(2S)-2-methyl-3-[(1-phenyl-1H-tetrazol-5-yl)sulfonyl]propyl}oxiran-2-yl)pentane-3-ol(P34)

This reaction was performed with reference to the literature (Wang, Z.,-X.; Tu, Y.; Frohn, M.; Zhang, J.-R.; Shi, Y., J. Am. Chem. Soc., 1997,119, 11224-11235. Wang, Z., -X.; Tu, Y.; Frohn, M.; Zhang, J.-R.; Shi,Y., J. Org. Chem., 1997, 62, 2328-2329.).

(3S,4S,5E,8S)-4,8-dimethyl-9-[(1-phenyl-1H-tetrazol-5-yl)sulfonyl]non-5-en-3-ol(190 mg, 0.50 mmol) was dissolved in acetonitrile (7.5 ml) and 0.05Msodium tetraborate decahydrate −0.4 mM disodiumethylenediaminetetraacetate salt solution (5 ml).1,2:4,5-di-O-isopropylidene-D-erythro-2,3-hexodiuro-2,6-pyranose (450mg, 1.74 mmol) was added while ice cooling. Subsequently, a mixed powderof potassium carbonate (0.85 mg, 6.16 mmol) and oxone (1.27 g, 2.06mmol) was added at the same temperature over one hour. The reactionsolution was stirred at the same temperature for further one hour. Thereaction solution was diluted with ethyl acetate and washed with waterand brine. After the organic layer was dried over anhydrous sodiumsulfate and the drying agent was filtered off, the organic layer wasconcentrated under reduced pressure. The obtained residue was purifiedby silica gel column chromatography (Kanto, commercial name Silica Gel60N, 40-100 μm; heptane:ethyl acetate=3:1) to obtain the title compound(160 mg, 94% ee) as a colorless oil. The optical purity was determinedby HPLC using a chiral column (DAICEL, commercial name CHIRALCEL OD;n-hexane:isopropylalcohol=75:25).

This oil was crystallized from hexane at −78° C. and recrystallizedfurther from a mixed solvent of hexane-ethyl acetate and the titlecompound (140 mg, >99% ee) was obtained as a colorless prism. Inaddition,(2S)-2-((2R,3R)-3-{(2S)-2-methyl-3-[(1-phenyl-1H-tetrazol-5-yl)sulfonyl]propyl}oxiran-2-yl)pentan-3-one(P34b, 34 mg) was obtained as colorless oil, as a by-product of thisprocedure.

Compound P34: 400 MHz ¹H-NMR (CD₃OD) δ (ppm) 0.98 (d, J=7.1 Hz, 3H),0.99 (t, J=7.3 Hz, 3H), 1.26 (d, J=6.8 Hz, 3H), 1.29-1.37 (m, 1H),1.45-1.63 (m, 3H), 1.89 (ddd, J=4.6, 5.8, 14.3 Hz, 1H), 2.45-2.57 (m,1H), 2.69 (dd, J=2.2, 7.9 Hz, 1H), 2.86 (ddd, J=2.2, 4.6, 6.9 Hz, 1H),3.57 (dt, J=4.6, 8.8 Hz, 1H), 3.69 (dd, J=7.3, 14.8 Hz, 1H), 3.91 (dd,J=5.2, 14.8 Hz, 1H), 7.66-7.77 (m, 5H); 100 MHZ ¹³C-NMR (CD₃OD) δ (ppm)10.54, 10.87, 20.29, 28.32, 28.57, 39.26, 42.23, 56.63, 61.67, 62.23,75.15, 126.94, 130.58, 132.54, 134.54, 155.41; IR (KBr) 3336, 3245,2961, 2925, 2900, 2869, 1779, 1594, 1460, 1334, 1154, 1109, 1071, 981,955, 828, 765, 688, 634, 524, 456 cm⁻¹; HRMS C₁₈H₂₇N₄O₄S (M+H⁺) Calcd:395.1753, Found: 395.1721; [α]_(D) ²⁰+21.0 (c 1.00, CH₃OH)

Compound P34b: 400 MHz ¹H-NMR (CDCL₃) δ (ppm) 1.03 (t, J=7.3 Hz, 3H),1.12 (d, J=7.1 Hz, 3H), 1.28 (d, J=6.9 Hz, 3H), 1.56 (ddd, J=7.4, 7.4,14.8 Hz, 1H), 1.96 (ddd, J=4.6, 5.8, 14.8 Hz, 1H), 2.31-2.45 (m, 1H),2.45-2.65 (m, 3H), 2.75-2.83 (m, 2H), 3.65 (dd, J=7.1, 14.6 Hz, 1H),3.93 (dd, J=5.3, 14.6 Hz, 1H), 7.56-7.71 (m, 5H); 100 MHZ ¹³C-NMR(CD₃OD) δ (ppm) 7.74, 12.93, 20.33, 28.30, 36.12, 39.00, 49.48, 56.62,60.05, 62.22, 127.03, 130.62, 132.60, 134.63, 155.47, 214.55; HRMS;C₁₈H₂₅N₄O₄S⁺ Calcd: 393.1597, Found: 393.1601 (M+H)⁺; [α]_(D) ²⁴+52.25(c 0.86, CH₃OH).

(2) Synthesis of5-({(2S)-3-[(2R,3R)-3-(1S,2S)-2-{[diethyl(isopropyl)silyl]oxy-1-methylbutyl)oxiran-2-yl}-2-methylpropyl}sulfonyl)-1-phenyl-1H-tetrazole(P³⁵)

Imidazole (345 mg, 5.07 mmol) and diethylisopropylsilyl chloride (418mg, 2.54 mmol) were added to a DMF (4 ml) solution of(2R,3S)-2-((2R,3R)-3-{(2S)-2-methyl-3-[(1-phenyl-1H-tetrazol-5-yl)sulfonyl]propyl}oxiran-2-yl)pentane-3-ol(200 mg, 0.507 mmol) at room temperature, and the mixture was stirred atthe same temperature for one hour. The reaction solution was dilutedwith ethyl acetate and washed with distilled water and brine. After theorganic layer was dried over anhydrous sodium sulfate and the dryingagent was filtered off, the organic layer was concentrated under reducedpressure. The obtained residue was purified by silica gel columnchromatography (Kanto, commercial name Silica Gel 60N, 40-100 μm;heptane:ethyl acetate=20:1→10:1) to obtain the title compound (265 mg)as a colorless oil.

400 MHz ¹H-NMR (CD₃OD) δ (ppm) 0.67-0.74 (m, 4H), 0.88 (t, J=7.2 Hz,3H), 0.95 (d, J=6.8 Hz, 3H), 0.90-0.98 (m, 1H), 1.04 (t, J=7.6 Hz, 3H),1.05 (t, J=7.6 Hz, 3H), 1.06 (d, J=5.6 Hz, 6H), 1.26 (d, J=6.8 Hz, 3H),1.32-1.43 (m, 1H), 1.52-1.63 (m, 3H), 1.92 (ddd, J=4.4, 6.0, 14.0 Hz,1H), 2.45-2.56 (m, 1H), 2.71 (dd, J=2.4, 7.6 Hz, 1H), 2.86 (ddd, J=2.4,4.4, 6.8 Hz, 1H), 3.70 (dd, J=7.6, 14.8 Hz, 1H), 3.80-3.86 (m, 1H), 3.90(dd, J=5.2, 14.8 Hz, 1H), 7.65-7.75 (m, 5H); 100 MHz ¹³C-NMR (CD₃OD) δ(ppm) 5.00, 5.17, 7.65, 10.02, 10.11, 14.40, 17.93, 20.31, 28.46, 28.64,39.40, 40.84, 57.28, 61.58, 62.28, 75.78, 126.99, 130.63, 132.58,134.71, 155.54; IR (neat) 2961, 2881, 1596, 1498, 1462, 1339, 1153,1014, 823, 763, 723, 633 cm⁻¹; HRMS C₂₅H₄₃N₄O₄SSi (M+H⁺) Calcd:523.2774, Found: 523.2766; [α]_(D) ²³+19.3 (c 1.09, CH₃OH)

[Step 17]

Synthesis of 6,7-O-[(S)-benzylidene]pladienolide A

(1) Synthesis of6,7-O-[(S)-benzylidene]-3-{[tert-butyl(dimethyl)silyl]oxy}-21-{[diethyl(isopropyl)silyl]oxy}-pladienolideA (36)

This reaction was performed with reference to the literature (Blakemore,P. R.; Cole, W. J.; Kociensky, P. J.; Morley, A., Synlett, 1998,26-28.).

THF distilled by using lithium aluminum hydride immediately before usewas used at this step.

15% toluene solution (0.317 ml) of potassium bis(trimethylsilyl)amidewas added dropwise to a THF (1 ml) solution of5-({(2S)-3-[(2R,3R)-3-(1S,2S)-2-{[diethyl(isopropyl)silyl]oxy-1-methylbutyl)oxiran-2-yl}-2-methylpropyl}sulfonyl)-1-phenyl-1H-tetrazole(62 mg, 0.119 mmol) at −78° C. under stirring and it was stirred at thesame temperature for 30 minutes. A THF (1 ml) solution of(2E)-3-((2S,3aS,4E,6S,7S,13aR)-11-{[tert-butyl(dimethyl)silyl]oxy}-6,13a-dimethyl-9-oxo-2-phenyl-6,7,9,10,11,12,13,13a-octahydro-3aH-[1,3]dioxolo[4,5-f]oxacyclododecin-7-yl)but-2-enal(94 mg, 0.179 mmol) was added dropwise to this reaction solution at −78°C. and stirred at the same temperature for one hour. The reactionsolution was warmed to room temperature and diluted with ethyl acetateafter addition of an appropriate amount of water and washed with brine.After the organic layer was dried over anhydrous sodium sulfate and thedrying agent was filtered off, the organic layer was concentrated underreduced pressure. The obtained residue was purified by silica gel columnchromatography (Kanto, Silica Gel 60N, 40-100 μm; heptane:ethylacetate=25:1-15:1) to obtain the title compound (65 mg) as a colorlessoil.

400 MHz ¹H-NMR (CD₃OD) δ (ppm) 0.15 (s, 3H), 0.17 (s, 3H), 0.66-0.75 (m,4H), 0.86 (t, J=7.6 Hz, 3H), 0.90 (d, J=6.8 Hz, 3H), 0.93 (d, J=6.8 Hz,3H), 0.95 (s, 9H), 0.94-1.03 (m, 1H), 1.05 (t, J=8.4 Hz, 6H), 1.07 (d,J=6.4 Hz, 6H), 1.12 (d, J=6.8 Hz, 3H), 1.24-1.35 (m, 1H), 1.38-1.50 (m,3H), 1.42 (s, 3H), 1.58 (dq, J=7.6, 14.4 Hz, 2H), 1.69-1.78 (m, 2H),1.78 (s, 3H), 1.96-2.05 (m, 1H), 2.28 (dd, J=10.2, 14.8 Hz, 1H),2.44-2.58 (m, 1H), 2.64-2.72 (m, 3H), 2.76 (dt, J=2.4, 6.0 Hz, 1H),3.80-3.86 (m, 1H), 4.05-4.13 (m, 1H), 4.28 (d, J=9.6 Hz, 1H), 4.96 (d,J=10.8 Hz, 1H), 5.50 (dd, J=9.6, 15.2 Hz, 1H), 5.68 (dd, J=9.6, 14.4 Hz,1H), 5.70 (dd, J=9.6, 15.2 Hz, 1H), 5.94 (s, 1H), 6.12 (d, 10.8 Hz, 1H),6.36 (dd, J=10.8, 14.4 Hz, 1H), 7.40-7.44 (m, 3H), 7.52-7.54 (m, 2H);100 MHz ¹³C-NMR (CD₃COCD₃) δ (ppm) −4.94, −4.85, 4.06, 4.24, 7.02, 7.05,9.44, 9.76, 11.17, 13.33, 16.70, 17.32, 17.92, 21.07, 22.61, 25.64,27.74, 31.69, 34.80, 35.51, 40.00, 40.08, 40.19, 44.08, 56.95, 60.92,71.89, 74.66, 82.34, 83.54, 85.35, 101.21, 124.75, 127.09, 128.24,129.11, 130.29, 130.76, 131.79, 137.63, 139.00, 141.19, 168.35; IR(neat)=2959, 1732, 1462, 1376, 1276, 1247, 1064, 1008, 971, 881, 836,775, 760, 722, 564, 536, 463 cm⁻¹; HRMS C₄₈H₈₀NaO₇Si₂ (M+Na⁺) Calcd:847.5340, Found: 847.5323.; [α]_(D) ²⁶+21.3 (c 1.09, CH₂Cl₂)

(2) Synthesis of 6,7-O-[(S)-benzylidene]pladienolide A (P37)

1M THF solution (3.80 ml, 696 μmol) of tetra-n-butylammonium fluoride(3.80 ml, 696 mmol) was added to a THF (3.80 ml) solution of6,7-O-[(S)-benzylidene]-3-{[tert-butyl(dimethyl)silyl]oxy}-21-{[diethyl(isopropyl)silyl]oxy}-pladienolideA (191 mg, 232 μmol) at room temperature. The reaction solution wasstirred at room temperature for one and a half hour. The reactionsolution was poured into a saturated ammonium chloride aqueous solution,which was then extracted with ethyl acetate. The organic layer waswashed with water and brine sequentially and dried over anhydrousmagnesium sulfate. After the drying agent was filtered off, the organiclayer was concentrated under reduced pressure. The obtained residue waspurified by silica gel column chromatography (Kanto Chemical, commercialname Silica gel 60N, granular, neutral, 0.040 mm-0.100 mm;n-heptane:ethyl acetate=2:1→1:1) to obtain the title compound (135 mg)as a colorless oil.

400 MHz, ¹H-NMR (CD₃OD) δ (ppm) 0.93 (d, J=7.2 Hz, 3H), 0.94 (d, J=7.2Hz, 3H), 0.98 (t, J=7.4 Hz, 3H), 1.13 (d, J=6.8 Hz, 3H), 1.18-1.29 (m,1H), 1.44 (s, 3H), 1.44-1.60 (m, 6H), 1.64-1.72 (m, 1H), 1.79 (d, J=1.2Hz, 3H), 2.00-2.10 (m, 1H), 2.33 (dd, J=9.8, 14.6 Hz, 1H), 2.46-2.56 (m,1H), 2.62-2.78 (m, 3H), 3.55 (dt, J=4.8, 8.0 Hz, 1H), 3.93-4.01 (m, 1H),4.28 (d, J=9.2 Hz, 1H), 4.97 (d, J=10.4 Hz, 1H), 5.49 (dd, J=9.6, 14.8Hz, 1H), 5.69 (dd, J=9.2, 14.8 Hz, 2H), 5.94 (s, 1H), 6.13 (brd, J=11.0Hz, 1H), 6.36 (dd, J=11.0, 14.8 Hz, 1H), 7.40-7.43 (m, 3H), 7.51-7.34(m, 2H); 100 MHz ¹³C-NMR (CD₃COCD₃) δ (ppm) 10.20, 10.30, 11.26, 16.48,20.87, 22.33, 27.88, 31.04, 34.09, 35.51, 39.80, 40.20, 41.53, 42.71,56.45, 61.31, 70.01, 73.62, 82.39, 83.66, 85.21, 101.20, 124.65, 127.09,128.30, 129.14, 130.49, 130.86, 131.74, 137.37, 139.11, 141.32, 169.85;IR (neat) 3502, 2965, 2931, 2874, 1711, 1456, 1377, 1246, 1221, 1178,1092, 1064, 971, 761 cm⁻¹; HRMS C₃₅H₅₀NaO₇ (M+Na⁺) Calcd: 605.3454,Found: 605.3449; [α]_(D) ²⁴−13.0 (c 1.05, CH₂Cl₂)

[Step 18]

Synthesis of Pladienolide A

(1) Synthesis of 6,7-O-[(S)-benzylidene]-3,21-bis(dichloroacetyl)pladienolide A (P38)

Triethylamine (686 μl, 4.92 mmol), dichloroacetic anhydride (375 μl,2.46 mmol) and 4-dimethylaminopyridine (15 mg, 123 μmol) were added toanhydrous dichloromethane (7.20 ml) solution of6,7-O-[(S)-benzylidene]pladienolide A (144 mg, 2.46 mmol) under stirringat 0° C. After the reaction solution was stirred at 0° C. for 40minutes, 4-dimethylaminopyridine (15 mg, 123 μmol) was added thereto andthe mixture was stirred at 0° C. for further one and a half hour. Thereaction solution was poured into water, which was then extracted withethyl acetate. The organic layer was washed with water and brinesequentially and dried over anhydrous magnesium sulfate. After thedrying agent was filtered off, the organic layer was concentrated underreduced pressure. The obtained residue was purified by silica gel columnchromatography (Merck, commercial name Silica Gel 60, 230-400 mesh;n-heptane:ethyl acetate=8:1→6:1→4:1) to obtain the title compound (198mg) as a yellow oil.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.88 (d, J=6.8 Hz, 3H), 0.93 (t, J=7.6Hz, 3H), 0.95 (d, J=6.8 Hz, 3H), 1.07 (d, J=6.8 Hz, 3H), 1.41 (s, 3H),1.46-1.60 (m, 3H), 1.62-1.68 (m, 1H), 1.68-1.81 (m, 4H), 1.73 (s, 3H),1,94-2.05 (m, 1H), 2.40-2.56 (m, 3H), 2.56-2.78 (m, 2H), 2.83 (dd,J=4.2, 15.0 Hz, 1H), 4.21 (d, J=9.2 Hz, 1H), 4.92 (d, J=10.4 Hz, 1H),5.02-5.08 (m, 1H), 5.12-5.22 (m, 1H), 5.42 (dd, J=9.6, 15.2 Hz, 1H),5.62 (dd, J=8.4, 14.8 Hz, 1H), 5.67 (dd, J=9.6, 15.2 Hz, 1H), 5.92 (s,1H), 5.93 (s, 1H), 5.96 (s, 1H), 6.09 (d, J=10.8 Hz, 1H), 6.24 (dd,J=10.8, 14.8 Hz, 1H), 7.37-7.42 (m, 3H), 7.49-7.52 (m, 2H); 100 MHz¹³C-NMR (CDCl₃) δ (ppm) 9.62, 10.96, 11.73, 16.48, 21.04, 22.62, 24.79,27.87, 32.99, 35.31, 38.87, 39.30, 39.55, 40.13, 56.41, 60.03, 64.14,64.58, 75.78, 80.54, 83.02, 83.19, 85.06, 101.26, 124.45, 126.61,128.33, 129.31, 129.64, 130.86, 131.04, 137.44, 137.74, 141.03, 164.06,167.56; IR (neat) 3015, 2972, 2931, 2877, 1759, 1739, 1576, 1457, 1377,1281, 1248, 1171, 1092, 1066, 1004, 968, 911, 816, 758, 699 cm⁻¹; HRMSC₃₉H₅₀Cl₄O₉Na (M+Na⁺) Calcd: 825.2107, Found: 825.2120; [α]_(D) ²³+15.0(c 1.02, CH₂Cl₂)

(2) Synthesis of 3,21-bis(dichloroacetyl) pladienolide A (24)

Pyridinium p-toluenesulfonate (70.5 mg, 280 μmol) was added to amethanol (4.48 ml) solution of6,7-O-[(S)-benzylidene]-3,21-bis(dichloroacetyl) pladienolide A (112 mg,140 mol) at room temperature. The reaction solution was stirred at roomtemperature for 46 hours. The reaction solution was poured into brine,which was then extracted with ethyl acetate. The organic layer waswashed with water and brine sequentially and dried over anhydrousmagnesium sulfate. After the drying agent was filtered off, the organiclayer was concentrated under reduced pressure. The obtained residue waspurified by silica gel column chromatography (Merck, commercial nameSilica Gel 60, 230-400 mesh; n-heptane:ethylacetate=8:1→6:1→3:1→3:2→1:1) to obtained title compound (32.2 mg) as acolorless oil while 6,7-O-[(S)-benzylidene]-3,21-bis(dichloroacetyl)pladienolide A (55.9 mg) was recovered.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.90 (d, J=7.2 Hz, 3H), 0.93 (t, J=7.2Hz, 3H), 0.96 (d, J=7.2 Hz, 3H), 1.06 (d, J=6.8 Hz, 3H), 1.32 (s, 3H),1.43-1.62 (m, 5H), 1.63-1.69 (m, 2H), 1.72 (s, 3H), 1.72-1.79 (m, 2H),2.00 (brs, 1H), 2.41-2.48 (m, 1H), 2.51-2.58 (m, 3H), 2.61 (dd, J=3.2,15.2 Hz, 1H), 2.66-2.70 (m, 1H), 2.74 (dd, J=4.0, 15.2 Hz, 1H), 3.80(dd, J=2.6, 9.8 Hz, 1H), 4.93-4.97 (m, 1H), 5.01-5.05 (m, 1H), 5.05 (d,J=10.4 Hz, 1H), 5.47 (dd, J=9.8, 15.3 Hz, 1H), 5.61 (dd, J=8.0, 15.1 Hz,1H), 5.74 (dd, J=9.6, 15.3 Hz, 1H), 5.97 (s, 1H), 5.98 (s, 1H), 6.07(brd, J=10.4 Hz, 1H), 6.24 (dd, J=10.4, 15.1 Hz, 1H); 100 MHz ¹³C-NMR(CDCl₃) δ (ppm) 9.68, 11.00, 11.77, 16.51, 21.08, 24.60, 24.83, 25.61,34.76, 35.35, 36.17, 39.34, 39.61, 40.47, 56.48, 60.07, 64.27, 64.61,73.28, 74.59, 80.60, 83.08, 124.52, 129.71, 130.86, 131.11, 137.79,141.06, 163.91, 164.17, 168.05; IR (neat) 3526, 2971, 2929, 1759, 1745,1456, 1377, 1281, 1217, 1171, 1092, 1008, 964, 927, 816, 757 cm⁻¹; HRMSC₃₂H₄₆Cl₄NaO₉ (M+Na⁺) Calcd: 737.1794, Found: 737.1778; [α]_(D) ²⁷+22.5(c 1.52, CHCl₃)

(3) Synthesis of Pladienolide A

Potassium carbonate (13.5 mg, 97.5 μmol) was added to a methanol (2.50ml) solution of 3,21-bis(dichloroacetyl) pladienolide A (69.9 mg, 97.5μmol) at room temperature. After the reaction solution was stirred atroom temperature for 20 minutes, it was poured into brine, which wasthen extracted with ethyl acetate. After the organic layer was driedover anhydrous sodium sulfate and the drying agent was filtered off, theorganic layer was concentrated under reduced pressure. The obtainedresidue was purified by silica gel column chromatography (KantoChemical, commercial name Silica gel 60N, granular, neutral, 0.040mm-0.100 mm; n-heptane:acetone=3:2→1:1) to obtain the title compound(46.2 mg) as a colorless oil. This was obtained as a white amorphous bypurifying by HPLC (Shiseido, commercial name CAPCELL PAK C18SG120;acetonitrile:water=37:63) followed by performing lyophilization.

600 MHz ¹H-NMR (CD₃OD) δ (ppm) 0.95 (d, J=7.0 Hz, 3H), 0.95 (d, J=6.7Hz, 3H), 0.98 (t, J=7.4 Hz, 3H), 1.13 (d, J=6.8 Hz, 3H), 1.24 (m, 1H),1.31 (s, 3H), 1.38 (m, 2H), 1.50 (m, 1H), 1.53 (m, 2H), 1.57 (m, 1H),1.61 (m, 1H), 1.68 (ddd, J=5.5, 5.9, 14.0 Hz, 1H), 1.79 (brs, 3H), 2.52(m, 1H), 2.56 (m, 2H), 2.61 (ddq, J=7.0, 9.8, 10.7 Hz, 1H), 2.70 (dd,J=2.2, 8.2 Hz, 1H), 2.77 (ddd, J=2.2, 5.9, 5.9 Hz, 1H), 3.56 (ddd,J=4.5, 4.5, 8.7 Hz, 1H), 3.74 (d, J=9.8 Hz, 1H), 3.81 (m, 1H), 5.07 (d,J=10.7 Hz, 1H), 5.42 (dd, J=9.8, 15.0 Hz, 1H), 5.70 (dd, J=8.4, 15.0 Hz,1H), 5.76 (dd, J=9.8, 15.0 Hz, 1H), 6.13 (brd, J=10.8 Hz, 1H), 6.37 (dd,J=10.8, 15.0 Hz, 1H); 150 MHz ¹³C-NMR (CD₃OD) δ (ppm) 10.8, 10.9, 11.9,17.1, 21.7, 24.4, 28.6, 30.5, 36.7, 37.6, 40.1, 40.7, 1.8, 42.8, 58.5,63.0, 70.7, 74.7, 75.3, 78.2, 84.4, 125.9, 131.6, 132.1, 132.6, 137.7,142.3, 171.9; IR (KBr) 3403, 2966, 2935, 2876, 1708, 1458, 1371, 1256,1177, 1060, 1021, 977, 903, 789 cm⁻¹; HRMS C₂₈H₄₆NaO₇ (M+Na⁺) Calcd:517.3141, Found: 517.3134; [α]_(D) ²⁷−1.54 (c 1.02, MeOH)

[Step 19]

Synthesis of Pladienolide B

Triethylamine (17.2 μl, 123 μmol), acetic anhydride (5.83 μl, 616 μmol)and 4-dimethylaminopyridine (1.50 mg, 12.3 μmol) were added to anhydrousdichloromethane (2.00 ml) solution of pladienolide A (30.5 mg, 61.6μmol) under stirring at 0° C. After the reaction solution was stirred at0° C. for one hour, it was poured into a saturated sodium hydrogencarbonate aqueous solution, which was then extracted with ethyl acetate.After the organic layer was washed with brine, it was dried overanhydrous magnesium sulfate. After the drying agent was filtered off,the organic layer was concentrated under reduced pressure. The obtainedresidue was purified by silica gel column chromatography (KantoChemical, commercial name Silica gel 60N, granular, neutral, 0.040mm-0.100 mm; n-heptane:acetone=3:1→2:1) to obtain the title compound(27.2 mg) as a colorless oil. This was obtained as a white amorphous bypurifying by HPLC (Shiseido, commercial name CAPCELL PAK C18SG120;acetonitrile:water=45:55) followed by performing lyophilization.

600 MHz ¹H-NMR (CD₃OD) δ (ppm) 0.93 (d, J=6.8 Hz, 3H), 0.95 (d, J=7.0Hz, 3H), 0.98 (t, J=7.4 Hz, 3H), 1.13 (d, J=6.8 Hz, 3H), 1.23 (s, 3H),1.24 (m, 1H), 1.41 (m, 1H), 1.42 (m, 1H), 1.51 (m, 1H), 1.53 (m, 2H),1.61 (m, 1H), 1.67 (m, 1H), 1.68 (m, 1H), 1.79 (brs, 3H), 2.10 (s, 3H),2.52 (m, 1H), 2.57 (m, 2H), 2.61 (ddq, J=6.8, 9.9, 10.1 Hz, 1H), 2.70(dd, J=2.2, 8.2 Hz, 1H), 2.77 (ddd, J=2.2, 5.9, 5.9 Hz, 1H), 3.55 (ddd,J=4.5, 4.5, 8.7 Hz, 1H), 3.82 (m, 1H), 5.09 (d, J=9.8 Hz, 1H), 5.09 (d,J=10.1 Hz, 1H), 5.61 (dd, J=9.9, 15.2 Hz, 1H), 5.70 (dd, J=8.4, 15.0 Hz,1H), 5.74 (dd, J=9.8, 15.2 Hz, 1H), 6.13 (brd, J=10.8 Hz, 1H), 6.37 (dd,J=10.8, 15.0 Hz, 1H); 150 MHz ¹³C-NMR (CD₃OD) δ (ppm) 10.8, 10.9, 11.9,16.9, 21.1, 21.7, 24.2, 28.6, 30.4, 31.1, 36.7, 37.5, 40.1, 40.7, 41.7,42.8, 58.5, 63.0, 70.4, 74.1, 75.3, 80.3, 84.3, 125.8, 127.0, 132.2,132.4, 141.6, 142.3, 171.8, 172.2; IR (KBr) 3447, 2966, 2935, 2875,1735, 1720, 1458, 1372, 1244, 1175, 1022, 978, 910, 551, 478 cm⁻¹; HRMSC₃₀H₄₈NaO₈ (M+Na⁺) Calcd: 559.3247, Found: 559.3227; [α]_(D) ²⁷+7.90 (c1.10, MeOH)

Synthesis of a Compound Analogous to Pladienolide B

Synthesis of 21-{[tert-butyl (dimethyl)silyl]oxy}pladienolide B (R2)

This reaction is performed with reference to the literature (Grubbs, R.H. “Hand book of Metathesis”, Wiley-VCH, 2003, v. 2, p 246-292).

(i)(2S,3S,4E,6S,7R,10R)-7,10-dihydroxy-3,7-dimethyl-2-[(1E)-1-methylbuta-1,3-dien-1-yl]-12-oxooxacyclododec-4-en-6-ylacetate (11.0 mg, 30.0 μmol) andtert-butyl[((1S,2S)-1-ethyl-2{(2R,3R)-3-[(2S)-2-methylbut-3-en-1-yl]oxiran-2-yl}propyl)oxy]dimethylsilane(24.0 mg 76.8 μmol) were dissolved in anhydrous dichloromethane underargon atmosphere and the second generation Grubbs catalyst;[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(phenylmethylene)-tricyclohexylphosphine)rutheniumwas added. The reaction solution was heated to reflux for three hours.After the reaction solution was cooled to room temperature, it wasconcentrated under reduced pressure. The obtained residue was purifiedby silica gel column chromatography (Kanto, commercial name Silica Gel60N, 40-100 μm; heptane:ethyl acetate=1:1) to obtain the title compound(5.1 mg) as a colorless amorphous.

400 MHz ¹H-NMR (CD₃OD) δ (ppm) 0.12 (s, 6H), 0.86 (t J=7.6 Hz, 3H), 0.90(d, J=7.2 Hz, 3H), 0.92 (d, J=7.6 Hz, 3H), 0.95 (s, 9H), 1.12 (d, J=6.8Hz, 3H), 1.23 (s, 3H), 1.22-1.78 (m, 9H), 1.78 (s, 3H), 2.10 (s, 3H),2.46-2.64 (m, 4H), 2.66 (dd, J=2.4, 8.8 Hz, 1H), 2.76 (dt, J=2.4, 6.0Hz, 1H), 3.78-3.86 (m, 2H), 5.00-5.14 (m, 2H), 5.52-5.80 (m, 3H), 6.12(brd, J=10.8 Hz, 1H), 6.36 (dd, J=10.8, 15.2 Hz, 1H); LRMS C₃₆H₆₂NaO₈Si(M+Na⁺) Calcd: 673.41, Found: 673.26.

[Total Synthesis of Pladienolide D]

[Step D1]

Methyl-(2E)-3-methyl-5-[(1-phenyl-1-H-tetrazol-5-yl)sulfonyl]pent-2-enoateandethyl-(2E)-3-methyl-5-[(1-phenyl-1-H-tetrazol-5-yl)thio]pent-2-enoate

(1-1) Synthesis ofmethyl-(2E)-3-methyl-5-[(1-phenyl-1-H-tetrazol-5-yl)thio]pent-2-enoate(Q1a)

5-Mercapto-1-phenyltetrazole (2.62 g, 14.60 mmol), triphenylphosphine(3.83 g, 14.60 mmol) and diisopropyl azodicarboxylate (95%, 2.96 g,14.60 mmol) were added to a THF (36.8 ml) solution ofmethyl-(2E)-5-hydroxy-3-methylpent-2-enoate (1.16 g, 7.31 mmol) whileice cooling. The reaction solution was warmed to room temperature andstirred for two hours. After the reaction solution was diluted withethyl acetate, it was washed with distilled water and brine. After theorganic layer was dried over anhydrous sodium sulfate and the dryingagent was filtered off, the organic layer was concentrated under reducedpressure. The obtained residue was purified by silica gel columnchromatography (Merck, commercial name Silica Gel 60, 230-400 mesh;hexane:ethyl acetate=5:1) to obtain the title compound (2.33 g) as awhite solid.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 2.22 (d, J=1.2 Hz, 3H), 2.70 (t, J=7.2Hz, 2H), 3.54 (t, J=7.2 Hz, 2H), 3.70 (s, 3H), 5.70-5.74 (q, J=1.2 Hz,1H), 7.52-7.59 (m, 5H); 100 MHz ¹³C-NMR (CDCl₃) δ (ppm) 18.41, 30.75,39.68, 50.94, 117.30, 123.73, 129.81, 130.18, 133.46, 153.79, 155.84,166.49; IR (KBr)=3071, 2991, 2948, 1707, 1648, 1593, 1499, 1437, 1412,1380, 1226, 1153, 1057, 877, 761, 693, 559, 485, 407 cm⁻¹; HRMSC₁₄H₁₆AgN₄O₂S (M+Ag⁺) Calcd: 411.0045, Found: 411.0073.

(1-2) Synthesis ofethyl-(2E)-3-methyl-5-[(1-phenyl-1-H-tetrazol-5-yl)thio]pent-2-enoate(Q1b)

This reaction was performed with reference to the literature (Mitsunobu,O., Synthesis, 1981, 1-28.). 5-Mercapto-1-phenyltetrazole (16.30 g,91.20 mmol), triphenylphosphine (27.30 g, 104 mmol) and diisopropylazodicarboxylate (95%, 21.10 g, 104 mmol) were added to a THF (410 ml)solution of ethyl-(2E)-5-hydroxy-3-methylpent-2-enoate (13.80 g, 86.90mmol) while ice cooling. The reaction solution was warmed to roomtemperature and stirred for four hours. After the reaction solution wasdiluted with ethyl acetate, it was washed with distilled water andbrine. After the organic layer was dried over anhydrous sodium sulfateand the drying agent was filtered off, the organic layer wasconcentrated under reduced pressure. The obtained residue was purifiedby silica gel column chromatography (Merck, commercial name Silica Gel60, 230-400 mesh; hexane:ethyl acetate=5:1) to obtain the title compound(27.16 g) as a white solid.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 1.26 (t, J=7.2 Hz, 3H), 2.22 (s, 3H),2.70 (t, J=7.2 Hz, 2H), 3.54 (t, J=7.2 Hz, 2H), 4.15 (q, J=7.2 Hz, 2H),5.72 (brs, 1H), 7.54-7.59 (m, 5H); 100 MHz ¹³C-NMR (CDCl₃) δ (ppm)14.05, 18.22, 30.59, 39.53, 59.48, 117.58, 123.57, 129.61, 129.98,133.33, 153.61, 155.24, 165.96; IR (KBr)=3064, 2981, 2939, 2903, 2358,2341, 1713, 1650, 1597, 1500, 1386, 1278, 1222, 1145, 1054, 763, 694cm⁻¹; HRMS C₁₅H₁₈AgN₄O₂S (M+Ag⁺) Calcd: 425.0201, Found: 425.0170.

(2-1) Synthesis ofmethyl-(2E)-3-methyl-5-[(1-phenyl-1-H-tetrazol-5-yl)sulfonyl]pent-2-enoate(Q2a)

This reaction was performed with reference to the literature (Shultz, H.S.; Freyermuth, H. B.; Buc, S. R., J. Org. Chem., 1963, 28(4),1140-1140.).

About 30% hydrogen peroxide solution (26.7 ml, 235 mmol) solution ofhexaammonium heptamolybdate tetrahydrate (2.91 mg, 2.35 mmol) was addedto an ethanol (150 ml) solution ofmethyl-(2E)-3-methyl-5-[(1-phenyl-1-H-tetrazol-5-yl)thio]pent-2-enoate(7.53 g, 24.70 mmol) at room temperature. After stirred for at the sametemperature for 12 hours, the reaction solution was diluted with ethylacetate and washed with distilled water and brine. After the organiclayer was dried over anhydrous sodium sulfate and the drying agent wasfiltered off, the organic layer was concentrated under reduced pressure.The obtained residue was purified by silica gel column chromatography(Merck, commercial name Silica Gel 60, 230-400 mesh; hexane:ethylacetate=4:1) to obtain the title compound (8.24 g) as a white solid.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 2.24 (d, J=1.2 Hz, 3H), 2.79-2.83 (m,2H), 3.71 (s, 3H), 3.88-3.92 (m, 2H), 5.77-5.78 (q, J=1.2 Hz, 1H),7.60-7.71 (m, 5H); 100 MHz ¹³C-NMR (CDCl₃) δ (ppm) 18.25, 32.44, 50.90,53.59, 117.67, 124.83, 129.51, 131.30, 132.66, 152.97, 165.99; IR(KBr)=3102, 3075, 2952, 2913, 1703, 1651, 1495, 1439, 1346, 1238, 1160,1047, 998, 923, 882, 764, 689, 593, 550, 507, 456, 420 cm⁻¹; HRMSC₁₄H₁₆AgN₄O₄S (M+Ag⁺) Calcd: 442.9943, Found: 442.9929.

(2-2) Synthesis ofethyl-(2E)-3-methyl-5-[(1-phenyl-1-H-tetrazol-5-yl)sulfonyl]pent-2-enoate(Q2b)

This reaction was performed with reference to the literature (Shultz, H.S.; Freyermuth, H. B.; Buc, S. R., J. Org. Chem., 1963, 28(4),1140-1140.).

About 30% hydrogen peroxide solution (47.4 ml, 418.0 mmol) solution ofhexaammonium heptamolybdate tetrahydrate (542 mg, 0.44 mmol) was addedto an ethanol (200 ml) solution ofethyl-(2E)-3-methyl-5-[(1-phenyl-1-H-tetrazol-5-yl)thio]pent-2-enoate(13.31 g, 41.80 mmol) at room temperature. After stirred for at the sametemperature for 12 hours, the reaction solution was diluted with ethylacetate and washed with distilled water and brine. After the organiclayer was dried over anhydrous sodium sulfate and the drying agent wasfiltered off, the organic layer was concentrated under reduced pressure.The obtained residue was purified by silica gel column chromatography(Merck, commercial name Silica Gel 60, 230-400 mesh; hexane:ethylacetate=4:1) to obtain the title compound (12.65 g) as a white solid.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 1.29 (t, J=7.2 Hz, 3H), 2.24 (d, J=1.2Hz, 3H), 2.78-2.82 (m, 2H), 3.88-3.92 (m, 2H), 4.17 (q, J=7.2 Hz, 2H),5.77 (brs, 1H), 7.59-7.71 (m, 5H); 100 MHz ¹³C-NMR (CDCl₃) δ (ppm)14.22, 18.51, 32.72, 53.93, 59.97, 118.44, 124.95, 129.77, 131.55,132.88, 152.56, 153.19, 165.81; IR (KBr)=3077, 3008, 2991, 2906, 1698,1660, 1495, 1342, 1235, 1158, 1045, 877, 764, 689, 589, 550, 508, 454cm⁻¹; HRMS C₁₅H₁₈AgN₄O₄S (M+Ag⁺) Calcd: 457.0100, Found: 457.0133.

[Step D2]

Synthesis of(2E,5E,7S,8S)-3,7-dimethyl-8-[(triethylsilyl)oxy]deca-2,5-dien-1-ol

(1-1) Synthesis ofmethyl-(2E,5E,7S,8S)-3,7-dimethyl-8-[(triethylsilyl)oxy]deca-2,5-dienoate(Q3a)

This reaction was performed with reference to the literature (Blakemore,P. R.; Cole, W. J.; Kociensky, P. J.; Morley, A., Synlett, 1998,26-28.).

THF used at this step was distilled by using lithium aluminum hydride.In addition, DME was distilled by using calcium hydride.

0.5M Potassium bis(trimethylsilyl)amide toluene solution (8.48 ml, 4.24mmol) was added dropwise to a DME (40 ml) solution ofmethyl-(2E)-3-methyl-5-[(1-phenyl-1-H-tetrazol-5-yl)sulfonyl]pent-2-enoate(1.19 g, 3.53 mmol) at −60° C. and the reaction solution was stirred atthe same temperature for 30 minutes. Subsequently a THF (5 ml) solutionof (2R,3S)-2-methyl-3-[(triethylsilyl)oxy]pentanal (1.63 g, 7.06 mmol)was added dropwise at −78° C. and the reaction solution was stirred fortwo hours. After the reaction solution was warmed to room temperature,distilled water was added. The reaction solution was diluted with ethylacetate and washed with brine. After the organic layer was dried overanhydrous sodium sulfate and the drying agent was filtered off, theorganic layer was concentrated under reduced pressure. The obtainedresidue was purified by silica gel column chromatography (Merck,commercial name Silica Gel 60, 230-400 mesh; hexane:diethyl ether=50:1)to obtain the title compound (1.17 g) as a colorless oil. The titlecompound was determined to be a mixture of E:Z=17:1 by ¹H-NMR.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.60 (q, J=8.0 Hz, 6H), 0.87 (t, J=7.2Hz, 3H), 0.95 (t, J=8.0 Hz, 9H), 0.96 (d, J=6.8 Hz, 3H), 1.35-1.50 (m,2H), 2.14 (d, J=1.2 Hz, 3H), 2.25-2.31 (m, 1H), 2.82 (d, J=6.4 Hz, 2H),3.44-3.48 (m, 1H), 3.69 (s, 3H), 5.36 (ddd, J=6.8, 13.6, 15.6 Hz, 1H),5.51 (dd, J=7.6, 15.2 Hz, 1H), 5.68 (q, J=1.2 Hz, 1H); 100 MHz ¹³C-NMR(CDCl₃) δ (ppm) 5.04, 6.78, 9.35, 15.41, 18.57, 26.65, 41.50, 43.86,50.48, 77.18, 115.38, 124.80, 136.84, 158.86, 166.97; IR (neat)=2958,2879, 2352, 2330, 1723, 1651, 1435, 1219, 1147, 1013, 740 cm⁻¹; HRMSC₁₉H₃₆AgO₃Si (M+Ag⁺) Calcd: 447.1485, Found: 447.1461; [α]_(D) ²¹−22.1(c 1.10, CHCl₃).

(1-2) Synthesis ofethyl-(2E,5E,7S,8S)-3,7-dimethyl-8-[(triethylsilyl)oxy]deca-2,5-dienoate(Q3b)

This reaction was performed with reference to a document (Blakemore, P.R.; Cole, W. J.; Kociensky, P. J.; Morley, A., Synlett, 1998, 26-28.).

THF used at this step was distilled from lithium hydride aluminum. Inaddition, DME was distilled from calcium hydride.

0.5M Toluene solution (64.2 ml) of potassium bis(trimethylsilyl)amidewas added dropwise to a DME (280 ml) solution ofethyl-(2E)-3-methyl-5-[(1-phenyl-1-H-tetrazol-5-yl)sulfonyl]pent-2-enoate(9.0 g, 25.7 mmol) at −60° C. and the reaction solution was stirred atthe same temperature for 30 minutes. Subsequently a THF (50 ml) solutionof (2R,3S)-2-methyl-3-[(triethylsilyl)oxy]pentanal (12.0 g, 52.1 mmol)was added dropwise at −78° C. and the reaction solution was stirred forone hour. After the reaction solution was warmed to room temperature,distilled water was added. The reaction solution was diluted with ethylacetate and washed with brine. After the organic layer was dried overanhydrous sodium sulfate and the drying agent was filtered off, theorganic layer was concentrated under reduced pressure. The obtainedresidue was purified by silica gel column chromatography (Merck,commercial name Silica Gel 60, 230-400 mesh; hexane:diethyl ether=100:1)to obtain the title compound (8.92 g) as a colorless oil. The titlecompound was determined to be a mixture of E:Z=18:1 by ¹H-NMR.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.60 (q, J=8.0 Hz, 6H), 0.87 (t, J=7.2Hz, 3H), 0.95 (d, J=7.6 Hz, 3H), 0.96 (t, J=8.0 Hz, 9H), 1.27 (t, J=7.2Hz, 3H), 1.36-1.53 (m, 2H), 2.13 (d, J=1.2 Hz, 3H), 2.26-2.32 (m, 1H),2.81 (d, J=6.8 Hz, 2H), 3.44-3.48 (m, 1H), 4.14 (q, J=7.2 Hz, 2H), 5.36(ddd, J=6.8, 13.6, 15.2 Hz, 1H), 5.51 (dd, J=7.6, 15.2 Hz, 1H),5.67-5.68 (m, 1H); 100 MHz ¹³C-NMR (CDCl₃) δ (ppm) 5.05, 6.77, 9.35,14.11, 15.37, 18.51, 26.67, 41.50, 43.89, 59.18, 77.20, 115.85, 124.88,136.78, 158.38, 166.52; IR (neat)=2958, 2879, 2362, 2345, 1719, 1650,1460, 1218, 1144, 1050, 1013, 740 cm⁻¹; HRMS C₂₀H₃₈AgO₃Si (M+Ag⁺) Calcd:461.1641, Found: 461.1640; [α]_(D) ²²−19.0 (c 2.06, CHCl₃).

(2-1) Synthesis of(2E,5E,7S,8S)-3,7-dimethyl-8-[(triethylsilyl)oxy]deca-2,5-dien-1-ol (Q4)

1M toluene solution (1.76 ml, 1.76 mmol) of diisobutylaluminum hydridewas added dropwise to an toluene (6 ml) solution ofmethyl-(2E,5E,7S,8S)-3,7-dimethyl-8-[(triethylsilyl)oxy]deca-2,5-dienoate(0.22 g, 0.64 mmol) at −78° C. and the reaction solution was stirred atthe same temperature for 30 minutes. The reaction solution was dilutedwith diethyl ether, followed by addition of a saturated potassium sodiumtartrate tetrahydrate aqueous solution (1.0 ml) and was warmed to roomtemperature and stirred for two hours. After the organic layer was driedover anhydrous sodium sulfate and the drying agent was filtered off, theorganic layer was concentrated under reduced pressure. The obtainedresidue was purified by silica gel column chromatography (Merck,commercial name Silica Gel 60, 230-400 mesh; hexane:diethyl ether=8:1)to obtain the title compound (150 mg) as a colorless oil.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.60 (q, J=7.6 Hz, 6H), 0.87 (t, J=7.6Hz, 3H), 0.95 (d, J=14.0 Hz, 3H), 0.96 (t, J=7.6 Hz, 9H), 1.35-1.50 (m,2H), 1.65 (brs, 3H), 2.24-2.29 (m, 1H), 2.70 (d, J=6.4 Hz, 2H), 3.45(dd, J=5.6 Hz, 1H), 4.16 (d, J=7.2 Hz, 2H), 5.32-5.48 (m, 3H); 100 MHz¹³C-NMR (CDCl₃) δ (ppm) 5.08, 6.85, 9.37, 15.73, 16.08, 26.72, 41.50,42.82, 59.10 77.39, 123.82, 126.73, 135.25, 138.53; IR (neat)=3330,2959, 2875, 1671, 1459, 1419, 1009, 740 cm⁻¹; HRMS C₁₈H₃₆AgO₂Si (M+Ag⁺)Calcd: 419.1536, Found: 419.1572; [α]_(D) ²¹−23.4 (c 1.45, CHCl₃).

(2-2) Synthesis of(2E,5E,7S,8S)-3,7-dimethyl-8-[(triethylsilyl)oxy]deca-2,5-dien-1-ol (Q4)

1M toluene solution of diisobutylaluminum hydride (52.2 ml, 52.2 mmol)was added dropwise to an anhydrous diethyl ether (37.5 ml) solution ofethyl-(2E,5E,7S,8S)-3,7-dimethyl-8-[(triethylsilyl)oxy]deca-2,5-dienoate(8.92 g, 25.20 mmol) at −78° C. and the reaction solution was stirred atthe same temperature for one hour. Methanol (1 ml) and a saturatedpotassium sodium tartrate tetrahydrate aqueous solution (9.3 ml) wereadded to the reaction solution, and the mixture was warmed to roomtemperature and stirred for two hours. After the organic layer was driedover anhydrous sodium sulfate and the drying agent was filtered off, theorganic layer was concentrated under reduced pressure. The obtainedresidue was purified by silica gel column chromatography (Merck,commercial name Silica Gel 60, 230-400 mesh; hexane:diethyl ether=8:1)to obtain the title compound (3.84 g) as a colorless oil. ¹H-NMR data ofthe title compound obtained by (2-2) is completely identical to those ofthe title compound obtained by (2-1).

[Step D3]

Synthesis of{(2R,3R)-3-[(2E,4S,5S)-5-hydroxy-4-methylhept-2-en-1-yl]-3-methyloxiran-2-yl}methyl4-methylbenzenesulfonate

(1) Synthesis of(2R,3R)-3-methyl-3-{(2E,4S,5S)-4-methyl-5-[(triethylsilyl)oxy]hept-2-en-1-yl}oxiran-2-yl)methanol(Q5)

This reaction was performed with reference to the literature (Katsuki,T.; Sharpless, K. B., J. Am. Chem. Soc., 1980, 102, 5976-5978. Gao, Y.;Hanson, R. M. H.; Klunder, J. M.; Ko, S. Y.; Masamune, H.; Sharpless, K.B., ibid., 1987, 109, 5765-5780.).

(i) Distilled anhydrous dichloromethane (22 ml) was added to activatedmolecular sieves 4A powder (880 mg) under Ar atmosphere and the solutionwas cooled to −30° C. Subsequently, after diethyl (−)-tartrate (2.01 ml,11.7 mmol) and titanium tetraisopropoxide (2.32 ml, 7.89 mmol) wereadded to the reaction solution under stirring and the mixture wasstirred for five minutes, a dichloromethane (4 ml) solution of(2E,5E,7S,8S)-3,7-dimethyl-8-[(triethylsilyl)oxy]deca-2,5-dien-1-ol(1.76 g, 5.63 mmol) was added dropwise. After stirred for 30 minutes, 5Mtert-butyl hydroperoxide decane solution (2.26 ml, 11.20 mmol) was addeddropwise to the reaction solution over 15 minutes. Then the reactionsolution was stirred at the same temperature for further 1.5 hours.Distilled water (5 ml) was added to the reaction solution and it wasfiltered through celite. The filtrate was diluted with ethyl acetate andwashed with distilled water and brine. After the organic layer was driedover anhydrous sodium sulfate and the drying agent was filtered off, theorganic layer was concentrated under reduced pressure. The obtainedresidue was purified by silica gel column chromatography (Kanto,commercial name Silica Gel 60N, 40-100 μm; heptane:ethyl acetate=5:1) toobtain the title compound (1.69 g, 90% de) as a colorless oil. Themeasurement of optical purity of the title compound was performed byconverting the title compound to{(2R,3R)-3-[(2E,4S,5S)-5-hydroxy-4-methylhepta-2en-1-yl]-3-methyloxiran-2-yl}methyl4-methylbenzenesulfonate (Q7) and determining by HPLC using a chiralcolumn (DAICEL, commercial name CHIRALPAK AD-H,hexane:isopropylalcohol=90:10).

(ii) Distilled anhydrous dichloromethane (12 ml) was added to activatedmolecular sieves 4A powder (500 mg) under Ar atmosphere and the solutionwas cooled to −30° C. Subsequently, after isopropyl (−)-tartrate (0.202ml, 0.96 mmol) and titanium tetraisopropoxide (0.189 ml, 0.640 mmol)were added to the reaction solution under stirring and the mixture wasstirred for five minutes, a dichloromethane (3 ml) solution of(2E,5E,7S,8S)-3,7-dimethyl-8-[(triethylsilyl)oxy]deca-2,5-dien-1-ol (1.0g, 3.20 mmol) was added dropwise. After stirred for 30 minutes, 5Mtert-butyl hydroperoxide decane solution (1.28 ml, 6.36 mmol) was addeddropwise to the reaction solution over 15 minutes. Then the reactionsolution was stirred at the same temperature for further 2 hours.Distilled water (5 ml) was added to the reaction solution and it wasfiltered through celite. The filtrate was diluted with ethyl acetate andwashed with distilled water and brine. After the organic layer was driedover anhydrous sodium sulfate and the drying agent was filtered off, theorganic layer was concentrated under reduced pressure. The obtainedresidue was purified by silica gel column chromatography (Kanto,commercial name Silica Gel 60N, 40-100 μm; heptane:ethyl acetate=5:1) toobtain the title compound (925 mg, 82% de) as a colorless oil. Themeasurement of optical purity of the title compound was performed byconverting the title compound to{(2R,3R)-3-[(2E,4S,5S)-5-hydroxy-4-methylhepta-2en-1-yl]-3-methyloxiran-2-yl}methyl4-methylbenzenesulfonate (Q7) and determining by HPLC using a chiralcolumn (DAICEL, commercial name CHIRALPAK AD-H,hexane:isopropylalcohol=90:10).

400 MHz ¹H-NMR (CD₃OD) δ (ppm) 0.67 (q, J=8.0 Hz, 6H), 0.93 (t, J=7.6Hz, 3H), 1.02 (d, J=6.8 Hz, 3H), 1.03 (t, J=8.0 Hz, 9H), 1.29 (s, 3H),1.45-1.58 (m, 2H), 2.23-2.35 (m, 3H), 2.97 (dd, J=4.8, 6.4 Hz, 1H), 3.56(dt, J=5.6, 6.0 Hz, 1H), 3.63 (dd, J=6.0, 12.0 Hz, 1H), 3.75 (dd, J=4.8,12.0 Hz, 1H), 5.13 (dt, J=7.2, 15.6 Hz, 1H), 5.57 (dd, J=7.6, 15.6 Hz,1H); 100 MHz ¹³C-NMR (CD₃OD) δ (ppm) 6.14, 7.43, 9.85, 16.39, 17.05,27.91, 42.79, 42.99, 61.68, 61.72, 63.68, 78.70, 125.23, 138.18; IR(neat)=3422, 2959, 2877, 1459, 1239, 1103, 1016, 740 cm⁻¹; HRMSC₁₈H₃₆AgO₃Si (M+Ag⁺) Calcd: 435.1485, Found: 435.1492; [α]_(D) ²¹−15.1(c 2.14, MeOH) (90% de).

(2) Synthesis of(2R,3R)-3-methyl-3-{(2E,4S,5S)-4-methyl-5-[(triethylsilyl)oxy]hept-2-en-1-yl}oxiran-2-yl)methyl4-methylbenzenesulfonate (Q6)

Triethylamine (1.86 ml, 12.90 mmol), 4-dimethylaminopyridine (23.7 mg,0.19 mmol) and p-toluenesulphonyl chloride (493 mg, 2.59 mmol) wereadded to a dichloromethane (9 ml) solution of(2R,3R)-3-methyl-3-{(2E,4S,5S)-4-methyl-5-[(triethylsilyl)oxy]hept-2-en-1-yl}oxiran-2-yl)methanol(425 mg, 1.29 mmol, 90% de) at room temperature. The reaction solutionwas stirred at the same temperature for two hours. After the reactionsolution was diluted with ethyl acetate, it was washed with distilledwater and brine. After the organic layer was dried over anhydrous sodiumsulfate and the drying agent was filtered off, the organic layer wasconcentrated under reduced pressure. The obtained residue was purifiedby silica gel column chromatography (Kanto, commercial name Silica Gel60N, 40-100 μm; heptane:ethyl acetate=15:1) to obtain the title compound(641 mg, 90% de) as a colorless oil. The measurement of optical purityof the title compound was performed by converting the title compound to{(2R,3R)-3-[(2E,4S,5S)-5-hydroxy-4-methylhept-2-en-1-yl]-3-methyloxiran-2-yl}methyl4-methylbenzenesulfonate (Q7) and determining by HPLC using a chiralcolumn (DAICEL, commercial name CHIRALPAK AD-H,hexane:isopropylalcohol=90:10).

400 MHz ¹H-NMR (CD₃OD), δ (ppm) 0.66 (q, j=8.0 Hz, 6H), 0.91 (t, j=7.6Hz, 3H), 0.99 (d, j=7.2 Hz, 3H), 1.02 (t, J=8.0 Hz, 9H), 1.21 (s, 3H),1.40-1.56 (m, 2H), 2.15-2.35 (m, 3H), 2.50 (s, 3H), 3.02 (dd, J=4.8, 6.8Hz, 1H), 3.54 (dt, J=5.2, 6.0 Hz, 1H), 4.10 (dd, J=6.8, 11.2 Hz, 1H),4.27 (dd, J=4.8, 11.2 Hz, 1H), 5.37 (dt, J=8.0, 15.6 Hz, 1H), 5.53 (dd,J=8.0, 15.6 Hz, 1H), 7.50 (ddd, J=1.0, 2.0, 6.8 Hz, 2H), 7.85 (dd,J=1.6, 6.8 Hz, 2H); 100 MHz ¹³C-NMR (CD₃OD) δ (ppm) 6.11, 7.51, 9.90,16.44, 17.20, 21.73, 27.87, 42.11, 42.92, 59.45, 61.85, 70.33, 78.54,124.60, 128.97, 131.07, 134.19, 138.53, 146.44; [α]_(D) ²¹+2.70 (c 2.10,MeOH).

(3) Synthesis of{(2R,3R)-3-[(2E,4S,5S)-5-hydroxy-4-methylhept-2-en-1-yl]-3-methyloxiran-2-yl}methyl4-methylbenzenesulfonate (Q7)

(i) To a THF (2 ml) solution of(2R,3R)-3-methyl-3-{(2E,4S,5S)-4-methyl-5-[(triethylsilyl)oxy]hept-2-en-1-yl}oxiran-2-yl)methyl4-methylbenzenesulfonate (440 mg, 0.91 mmol, 90% de), 1M hydrochloricacid aqueous solution (0.1 ml) was added at room temperature and thereaction was stirred at the same temperature for one hour. After thereaction solution was diluted with ethyl acetate, it was washed withdistilled water and brine. After the organic layer was dried overanhydrous sodium sulfate and the drying agent was filtered off, theorganic layer was concentrated under reduced pressure. The obtainedresidue was purified by silica gel column chromatography (Kanto,commercial name Silica Gel 60N, 40-100 μm; heptane:ethyl acetate=2:1) toobtain the title compound (334 mg, 90% de) as a colorless oil. Theoptical purity was determined by HPLC using a chiral column (DAICEL,commercial name CHIRALPAK AD-H, hexane:isopropylalcohol=90:10).

(ii) The title compound({(2R,3R)-3-[(2E,4S,5S)-5-hydroxy-4-methylhept-2-en-1-yl]-3-methyloxiran-2-yl}methyl4-methylbenzenesulfonate) (850 mg, 82% de) was purified by HPLC using apreparative chiral column (DAICEL for fractionation, commercial nameCHIRALPAK AD, 20×250 mm, hexane:isopropylalcohol=90:10, flow rate 10mL/min) and the title compound (634 mg, >99% de) was obtained as acolorless oil. In addition, β-epoxy isomer thereof (38 mg, >99% de) wasobtained as a colorless oil.

Q7: 400 MHz ¹H-NMR (CD₃OD) δ (ppm) 0.97 (t, J=7.6 Hz, 3H), 1.04 (d,J=6.8 Hz, 3H), 1.21 (s, 3H), 1.29-1.38 (m, 1H), 1.53-1.63 (m, 1H),2.14-2.30 (m, 3H), 2.50 (s, 3H), 3.03 (dd, J=4.4, 6.8 Hz, 1H), 3.25-3.29(m, 1H), 4.10 (dd, J=6.8, 11.2 Hz, 1H), 4.28 (dd, J=4.4, 11.2 Hz, 1H),5.38 (dt, J=6.8, 15.6 Hz, 1H), 5.49 (dd, J=8.0, 15.6 Hz, 1H), 7.50 (dd,J=1.0, 8.4 Hz, 2H), 7.85 (ddd, J=2.0, 2.0, 8.4 Hz, 2H); 100 MHz ¹³C-NMR(CD₃OD) δ (ppm) 10.64, 16.58, 17.10, 21.62, 28.31, 41.96, 44.21, 59.51,62.01, 70.43, 77.63, 124.85, 129.00, 131.13, 134.15, 138.80, 146.65; IR(neat)=3386, 2976, 2958, 2924, 2502, 1933, 1599, 1454, 1359, 1173, 1095,967, 866, 782, 667, 554 cm⁻¹; HRMS C₁₉H₂₈NaO₅S (M+Na⁺) Calcd: 391.1555,Found: 391.1550; [α]_(D) ²⁰+2.82 (c1.04, MeOH).

β-epoxy isomer (isomer of Q7): 400 MHz ¹H-NMR (CD₃OD) δ (ppm) 0.98 (t,J=7.6 Hz, 3H), 1.04 (d, J=6.8 Hz, 3H), 1.21 (s, 3H), 1.27-1.42 (m, 1H),1.53-1.65 (m, 1H), 2.13-2.31 (m, 3H), 2.50 (s, 3H), 3.03 (dd, J=4.8, 6.8Hz, 1H), 3.23-3.31 (m, 1H), 4.10 (dd, J=6.8, 11.6 Hz, 1H), 4.28 (dd,J=4.8, 11.6 Hz, 1H), 5.38 (dt, J=7.2, 15.6 Hz, 1H), 5.49 (dd, J=8.0,15.6 Hz, 1H), 7.50 (dd, J=0.8, 8.0 Hz, 2H), 7.85 (m, 2H).

[Step D4]

Synthesis of(1R)-1,2-anhydro-3,5-dideoxy-1-[(1R,2S)-2-hydroxy-1-methylbutyl]-4-C-vinyl-D-erythro-pentitol

(1) Synthesis of5,6:8,9-dianhydro-1,2,4,7-tetradeoxy-4,8-dimethyl-10-O-[(4-methylphenyl)sulfonyl]-D-threo-D-galacto-decitol(Q8)

This reaction was performed with reference to the literature (Wang, Z.,-X.; Tu, Y.; Frohn, M.; Zhang, J.-R.; Shi, Y., J. Am. Chem. Soc., 1997,119, 11224-11235. Wang, Z., -X.; Tu, Y.; Frohn, M.; Zhang, J.-R.; Shi,Y., J. Org. Chem., 1997, 62, 2328-2329.).

(i){(2R,3R)-3-[(2E,4S,5S)-5-hydroxy-4-methylhept-2-en-1-yl]-3-methyloxiran-2-yl}methyl4-methylbenzenesulfonate (2.82 g, 7.64 mmol, 89% de) was dissolved inacetonitrile (80.8 ml) and 0.4 mM ethylenediaminetetraacetic aciddisodium salt solution (53.4 ml) of 0.05M sodium tetraboratedecahydrate.1,2:4,5-di-O-isopropylidene-D-erythro-2,3-hexodiuro-2,6-pyranose (1.97g, 7.64 mmol) was added while ice cooling. Subsequently, a mixed powderof potassium carbonate (12.70 g, 91.80 mmol) and oxone (14.10 g, 22.93mmol) were added at the same temperature over four hours. The reactionsolution was stirred at the same temperature for another hour. Thereaction solution was diluted with ethyl acetate and washed withdistilled water and brine. After the organic layer was dried overanhydrous sodium sulfate and the drying agent was filtered off, theorganic layer was concentrated under reduced pressure. The obtainedresidue was purified by silica gel column chromatography (Kanto,commercial name Silica Gel 60N, 40-100 μm; heptane:ethylacetate=3:1→2:1) to obtain the title compound (2.57 g, 81% de) as acolorless oil. The optical purity was determined by HPLC using a chiralcolumn (DAICEL, commercial name CHIRALPAK AD-H,hexane:isopropylalcohol=80:20). Subsequently, it was recrystallized witha mixed solvent of hexane-diethyl ether and the title compound (1.52 g)was obtained as a colorless prism crystal of a single diastereomer (>99%de).

(ii){(2R,3R)-3-[(2E,4S,5S)-5-hydroxy-4-methylhept-2-en-1-yl]-3-methyloxiran-2-yl}methyl4-methylbenzenesulfonate (700 mg, 1.90 mmol, >99% de) was dissolved inacetonitrile (20.1 ml) and 0.4 mM ethylenediaminetetraacetic aciddisodium salt solution (13.3 ml) of 0.05M sodium tetraboratedecahydrate.1,2:4,5-di-O-isopropylidene-D-erythro-2,3-hexodiuro-2,6-pyranose (489mg, 1.90 mmol) was added while ice cooling. Subsequently, a mixed powderof potassium carbonate (3.15 g, 22.8 mmol) and oxone (3.50 g, 5.71 mmol)were added at the same temperature over four hours. The reactionsolution was further stirred at the same temperature for one hour. Thereaction solution was diluted with ethyl acetate and washed withdistilled water and brine. After the organic layer was dried overanhydrous sodium sulfate and the drying agent was filtered off, theorganic layer was concentrated under reduced pressure. The obtainedresidue was purified by silica gel column chromatography (Kanto,commercial name Silica Gel 60N, 40-100 μn; heptane:ethylacetate=3:1→2:1) to obtain the title compound (331 mg, 88% de) as acolorless oil. The optical purity was determined by HPLC using a chiralcolumn (DAICEL, commercial name CHIRALPAK AD-H,hexane:isopropylalcohol=80:20).

400 MHz ¹H-NMR (CD₃OD) δ (ppm) 0.98 (d, j=7.2 Hz, 3H), 0.99 (t, J=7.6Hz, 3H), 1.27-1.36 (m, 1H), 1.32 (s, 3H), 1.48-1.61 (m, 2H), 1.67 (dd,J=6.8, 14.4 Hz, 1H), 1.80 (dd, J=4.8, 14.4 Hz, 1H), 2.50 (s, 3H), 2.71(dd, J=6.4, 8.0 Hz, 1H), 2.85-2.89 (m, 1H), 3.10 (dd, J=4.4, 6.4 Hz,1H), 3.57 (dt, J=4.8, 8.0 Hz, 1H), 4.12 (dd, J=6.4, 11.2 Hz, 1H), 4.31(dd, J=4.4, 11.6 Hz, 1H), 7.44 (dd, J=1.0, 8.0 Hz, 2H), 7.86 (dd, J=2.0,8.4 Hz, 1H), 7.87 (dd, J=2.0, 8.4 Hz, 1H); 100 MHz ¹³C-NMR (CD₃OD) δ(ppm) 10.55, 10.85, 17.41, 21.62, 28.55, 41.82, 42.36, 55.43, 60.18,60.63, 61.63, 70.17, 75.15, 129.00, 131.13, 134.04, 146.66; IR(KBr)=3389, 2976, 2957, 2873, 1923, 1598, 1454, 1359, 1188, 1173, 1099,969, 866, 815, 669, 557, 507 cm⁻¹; HRMS C₁₉H₂₈AgO₆S (M+Ag⁺) Calcd:491.0658, Found: 491.0638; [α]_(D) ²³+42.7 (c 1.00, MeOH) (>99% de).

(2) Synthesis of(1R)-1,2-anhydro-3,5-dideoxy-1-[(1R,2S)-2-hydroxy-1-methylbutyl]-4-C-vinyl-D-erythro-pentitol(Q9)

This reaction was performed with reference to the literature (Fujii, N.;Habashita, H.; Akaji, M.; Nakai, K.; Ibuka, T.; Fujiwara, M.; Tamamura,H.; Yamamoto, Y., J. Chem. Soc., Perkin Trans. 1, 1996, 865-866.).

5,6:8,9-dianhydro-1,2,4,7-tetradeoxy-4,8-dimethyl-10-O-[(4-methylphenyl)sulfonyl]-D-threo-D-galacto-decitol(350 mg, 0.91 mmol) was dissolved in a mixture of acetone (17.5 ml) andDMF (3.5 ml), followed by addition of potassium iodide (528 mg, 3.18mmol), and the mixture was heated to reflux for two hours. The reactionsolution was cooled to 0° C., followed by addition of4-(dimethylamino)phenyldiphenylphosphine (420 mg, 1.37 mmol) and iodine(100 mg, 0.45 mmol), and the mixture was stirred at the same temperaturefor 15 minutes. 5% Sodium hydrogen carbonate aqueous solution (3 ml) and5% carbonic acid sodium thiosulfate aqueous solution (2 ml) were addedto the reaction solution and it was stirred at the same temperature forfurther 10 minutes. The reaction solution was diluted with ethyl acetateand washed with distilled water and brine. After the organic layer wasdried over anhydrous sodium sulfate and the drying agent was filteredoff, the organic layer was concentrated under reduced pressure. Theobtained residue was purified by silica gel column chromatography(Kanto, commercial name Silica Gel 60N, 40-100 μm; heptane:ethylacetate=2.5:1) to obtain the title compound (186 mg) as a colorless oil.

400 MHz ¹H-NMR (CD₃OD) δ (ppm) 0.98 (d, J=7.2 Hz, 3H), 0.99 (t, J=7.6Hz, 3H), 1.29-1.37 (m, 1H), 1.36 (s, 3H), 1.48-1.61 (m, 2H), 1.73 (dd,J=6.0, 14.0 Hz, 1H), 1.80 (dd, J=6.0, 14.0 Hz, 1H), 2.72 (dd, J=2.4, 7.6Hz, 1H), 2.95 (dt, J=2.4, 6.0 Hz, 1H), 3.38 (s, 1H), 3.58 (dt, J=4.8,8.0 Hz, 1H), 5.08 (dd, J=1.6, 10.8 Hz, 1H), 5.29 (dd, J=1.6, 17.6 Hz,1H), 6.02 (dd, J=10.8, 17.6 Hz, 1H); 100 MHz ¹³C-NMR (CD₃OD) δ (ppm)10.28, 10.92, 27.88, 28.54, 42.39, 45.54, 55.74, 62.10, 73.17, 75.18,112.21, 146.27; IR (neat)=3418, 3088, 2970, 2935, 2879, 1647, 1455,1416, 925 cm⁻¹; HRMS C₁₂H₂₂O₃Ag (M+Ag⁺) Calcd: 321.0620, Found:321.0667; [α]_(D) ²⁴+13.5 (c 1.67, MeOH) (>99% de).

(2-1) Alternative synthesis of(1R)-1,2-anhydro-3,5-dideoxy-1-[(1R,2S)-2-hydroxy-1-methylbutyl]-4-C-vinyl-D-erythro-pentitol(Q9)

This reaction was performed with reference to the literature(Sarandeses, L. A.; Mourino, A.; Luche, J-L., J. Chem. Soc., Chem.Commun., 1991, 818-820.).

5,6:8,9-dianhydro-1,2,4,7-tetradeoxy-4,8-dimethyl-10-O-[(4-methylphenyl)sulfonyl]-D-threo-D-galacto-decitol(200 mg, 0.53 mmol) was dissolved in a mixed solvent of acetone (5 ml)and DMF (1 ml), followed by addition of potassium iodide (303 mg, 1.83mmol). The mixture was heated to reflux for two hours. The reactionsolution was cooled to room temperature. 5% sodium hydrogen carbonateaqueous solution (3 ml) and 5% carbonic acid sodium thiosulfate aqueoussolution (2 ml) were added to the reaction solution and it was stirredat the same temperature for further 10 minutes. The reaction solutionwas diluted with ethyl acetate and washed with distilled water andbrine. After the organic layer was dried over anhydrous sodium sulfate,the drying agent was filtered off and the organic layer was concentratedunder reduced pressure. The obtained crude product was used in the nextstep.

Zinc powder (102, 1.56 mmol) was dissolved in a mixed solution ofethanol (0.5 ml) and water (0.75 ml), followed by addition of copperiodide (99 mg, 0.52 mmol) and the mixed solution was treated withsonication for 5 minutes. An ethanolic solution of the above-obtainedcrude product (0.5 ml) was added to the reaction solution and it wastreated with sonication at same temperature for 1.5 hours. The reactionsolution was diluted with ethyl acetate and filtered through celite andwashed with distilled water and brine. The organic layer was dried overanhydrous sodium sulfate and the drying agent was filtered off, theorganic layer was concentrated under reduced pressure. The obtainedresidue was purified by silica gel column chromatography (Kanto,commercial name Silica Gel 60N, 40-100 μm; heptane:ethylacetate=2.5:1)to obtain the title compound (110 mg) as a colorless oil. ¹H-NMR data oftitle compound by this alternative method are completely identical tothose of the title compound obtained by the above (2).

[Step D5]

Synthesis of(2S,3S,4E,6S,7R,10R)-7,10-dihydroxy-3,7-dimethyl-2-[(1E)-1-methylbuta-1,3-dien-1-yl]-12-oxooxacyclododec-4-en-6-ylacetate and(2S,3S,4E,6S,7R,10R)-7-hydroxy-3,7-dimethyl-2-[(1E)-1-methylbuta-1,3-dien-1-yl]-12-oxo-10-[(triethylsilyl)oxy]oxacyclododec-4-en-6-ylacetate

(1) Synthesis of(2S,3aS,4E,6S,7S,11R,13aR)-11-{[tert-butyl(dimethyl)silyl]oxy}-6,13a-dimethyl-7-[(1E)-1-methylbuta-1,3-dien-1-yl]-2-phenyl-3a,6,7,10,11,12,13,13a-octahydro-9H-[1,3]dioxolo[4,5-f]oxacyclododecin-9-one(Q10)

This reaction was performed with reference to the literature (Pine, S.H.; Zahler, R.; Evans, D. A.; Grubbs, R. H., J. Am. Chem. Soc., 1980,3270-3272.).

(2S,3aS,4E,6S,7S,13aR)-11-{[tert-butyl(dimethyl)silyl]oxy}-7-[(E)-2-formyl-1-methyleth-1-en-1-yl]-6,13a-dimethyl-2-phenyl-3a,6,7,10,11,12,13,13a-octahydro-9H-[1,3]dioxolo[4,5-f]oxacyclododecin-9-one(50 mg, 0.09 mmol) was dissolved in a mixed solvent of THF (0.4 ml) andtoluene (2.8 ml), and the solution was cooled to −40° C. After additionof pyridine (0.38 ml, 4.73 mmol) and 0.5M toluene solution (1.14 ml,0.57 mmol) of Tebbe reagent(chlorobis(cyclopentadienyl)-(dimethylaluminum)-methylene titanium), themixture was stirred at the same temperature for 30 minutes. It waswarmed to room temperature and further stirred for one hour. Distilledwater (1 ml) was added to the reaction solution, and the reactionsolution was diluted with ethyl acetate and washed with distilled waterand brine sequentially, and after dried over anhydrous sodium sulfate,the drying agent was filtered off and the organic layer was concentratedunder reduced pressure. The obtained residue was purified by silica gelcolumn chromatography (Merck, commercial name Silica Gel 60, 230-400mesh; hexane:diethyl ether=4:1) to obtain the title compound (36.3 mg)as a colorless amorphous substance.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.07 (s, 3H), 0.09 (s, 3H), 0.88 (s, 9H),0.89 (d, J=6.0 Hz, 3H), 1.35-1.47 (m, 2H), 1.38 (s, 3H), 1.59-1.67 (m,1H), 1.74 (d, J=0.8 Hz, 3H), 2.01-2.09 (m, 1H), 2.31 (dd, J=10.4, 14.4Hz, 1H), 2.58 (dd, J=4.4, 14.4 Hz, 1H), 2.54-2.63 (m, 1H), 3.94-4.02 (m,1H), 4.19 (d, J=9.2 Hz, 1H), 4.97 (d, J=10.4 Hz, 1H), 5.18 (dd, J=1.6,17.6 Hz, 1H), 5.27 (dd, J=1.6, 17.2 Hz, 1H), 5.40 (dd, J=9.6, 15.2 Hz,1H), 5.64 (dd, J=9.6, 15.2 Hz, 1H), 5.91 (s, 1H), 6.13 (d, J=10.8 Hz,1H), 6.54 (dt, J=10.4, 17.2 Hz, 1H), 7.34-7.41 (m, 3H), 7.48-7.52 (m,2H); 100 MHz ¹³C-NMR (CDCl₃) δ (ppm) −4.63, −4.51, 11.63, 16.72, 17.93,22.82, 25.72, 31.59, 34.42, 40.24, 44.11, 71.62, 82.07, 83.61, 85.32,101.21, 118.83, 126.75, 128.32, 129.28, 129.57, 131.26, 131.87, 133.41,137.69, 137.76, 169.18; IR (KBr)=2957, 2931, 2857, 1735, 1459, 1375,1275, 1244, 1174, 1078, 1005, 975, 908, 877, 835, 775, 759, 698 cm⁻¹;HRMS C₃₁H₄₆AgO₅Si (M+Ag⁺) Calcd: 633:2165, Found: 633.2189; [α]_(D)²³+19.5 (c 0.90, CHCl₃).

(2) Synthesis of(4R,7R,8S,9E,11S,12S)-4,7,8-trihydroxy-7,11-dimethyl-12-[(1E)-1-methylbuta-1,3-dien-1-yl]-oxacyclododec-9-en-2-one(Q11)

Pyridinium p-toluene sulfonate (68.2 mg, 271 μmol) was added to amethanol (1.22 ml) solution of(2S,3aS,4E,6S,7S,11R,13aR)-11-{[tert-butyl(dimethyl)silyl]oxy}-6,13a-dimethyl-7-[(1E)-1-methylbuta-1,3-dien-1-yl]-2-phenyl-3a,6,7,10,11,12,13,13a-octahydro-9H-[1,3]dioxolo[4,5-f]oxacyclododecin-9-one(36.0 mg, 0.07 mmol) and the reaction solution was stirred at roomtemperature for four days. Distilled water (1 ml) was added to thereaction solution, and the reaction solution was diluted with ethylacetate and washed with distilled water and brine sequentially, andafter dried over anhydrous sodium sulfate, the drying agent was filteredoff and the organic layer was concentrated under reduced pressure. Theobtained residue was purified by silica gel column chromatography(Merck, commercial name Silica Gel 60, 230-400 mesh; hexane:diethylether=1:3) to obtain the title compound (18.3 mg) as a white solid.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.92 (d, J=6.9 Hz, 3H), 1.24-1.32 (m,1H), 1.31 (s, 3H), 1.36-1.72 (m, 3H), 1.76 (d, J=1.0 Hz, 3H), 2.51-2.65(m, 3H), 3.72-3.77 (m, 1H), 3.82 (d, J=9.6 Hz, 1H), 5.17 (d, J=10.8 Hz,1H), 5.20 (d, J=11.2 Hz, 1H), 5.28 (dd, J=1.6, 16.8 Hz, 1H), 5.43 (dd,J=10.0, 15.2 Hz, 1H), 5.73 (dd, J=10.0, 15.2 Hz, 1H), 6.12 (d, J=11.6Hz, 1H), 6.55 (dt, J=10.4, 16.8 Hz, 1H); 100 MHz ¹³C-NMR (CDCl₃) δ (ppm)11.75, 16.52, 24.45, 29.72, 35.57, 38.23, 40.66, 69.23, 73.39, 77.12,82.45, 119.17, 130.19, 131.61, 131.81, 132.94, 137.18, 172.16; IR(KBr)=3473, 3336, 2960, 2928, 2872, 1729, 1604, 1580, 1460, 1274, 1122,1028, 959, 743, 420 cm⁻¹; HRMS C₁₈H₂₈AgO₅ (M+Ag⁺) Calcd: 431.0988,Found: 431.0960; [α]_(D) ²²+7.01 (c 1.09, CHCl₃).

(3) Synthesis of(2S,3S,4E,6S,7R,10R)-7,10-dihydroxy-3,7-dimethyl-2-[(1E)-1-methylbuta-1,3-dien-1-yl]-12-oxooxacyclododec-4-en-6-ylacetate (Q12)

Triethylamine (35.6 μl, 246 μmol), acetic anhydride (11.6 μl, 123 mol)and 4-dimethylaminopyridine (3.0 mg, 24.6 μmol) were added to ananhydrous dichloromethane (2 ml) solution of(4R,7R,8S,9E,11S,12S)-4,7,8-trihydroxy-7,11-dimethyl-12-[(1E)-1-methylbuta-1,3-dien-1-yl]-oxacyclododec-9-en-2-one(40.0 mg, 123 μmol) at 0° C. and the reaction solution was stirred atthe same temperature for one hour. The reaction solution was poured intoa saturated sodium hydrogen carbonate aqueous solution, and the reactionsolution was diluted with ethyl acetate and washed with distilled waterand brine sequentially, and after dried over anhydrous sodium sulfate,the drying agent was filtered off and the organic layer was concentratedunder reduced pressure. The obtained residue was purified by silica gelcolumn chromatography (Merck, commercial name Silica Gel 60, 230-400mesh; hexane:diethyl ether=1:2) to obtain the title compound (42 mg) asa white solid.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.90 (d, J=6.8 Hz, 3H), 1.21 (s, 3H),1.25-1.42 (m, 2H), 1.51-1.74 (m, 2H), 1.75 (s, 3H), 2.09 (s, 3H), 2.16(s, 1H), 2.46-2.67 (m, 3H), 3.54 (d, J=11.2 Hz, 1H), 3.70-3.80 (m, 1H),5.09 (d, J=8.8 Hz, 1H), 5.17 (d, J=10.8 Hz, 1H), 5.18 (d, J=10.0 Hz,1H), 5.28 (d, J=16.8 Hz, 1H), 5.61 (dd, J=9.6, 15.2 Hz, 1H), 5.68 (dd,J=9.6, 15.2 Hz, 1H), 6.12 (d, J=10.8 Hz, 1H), 6.55 (dt, J=10.4, 16.8 Hz,1H); 100 MHz ¹³C-NMR (CDCl₃) δ (ppm) 11.75, 16.40, 21.24, 24.57, 29.75,35.15, 38.26, 40.71, 69.11, 73.35, 78.85, 82.39, 119.19, 125.57, 131.65,131.79, 132.88, 140.39, 169.61, 172.00; IR (KBr)=3513, 3404, 2970, 2948,2876, 1734, 1705, 1454, 1429, 1405, 1361, 1248, 1175, 1051, 1023, 968,921, 652, 550, 508 cm⁻¹; HRMS C₂₀H₃₀O₆Na (M+Na⁺) Calcd: 389.1940, Found:389.1927; [α]_(D) ²²+22.2 (c 1.21, CHCl₃).

(4) Synthesis of(2S,3S,4E,6S,7R,10R)-7-hydroxy-3,7-dimethyl-2-[(1E)-1-methylbuta-1,3-dien-1-yl]-12-oxo-10-[(triethylsilyl)oxy]oxacyclododec-4-en-6-ylacetate (Q13)

Triethylamine (260 μl, 1.80 mmol), 4-dimethylaminopyridine (4.26 mg,0.349 mmol) and chlorotriethylsilane (48 μl, 0.286 mmol) were added toan anhydrous dichloromethane (1.5 ml) solution of(2S,3S,4E,6S,7R,10R)-7,10-dihydroxy-3,7-dimethyl-2-[(1E)-1-methylbuta-1,3-dien-1-yl]-12-oxooxacyclododec-4-en-6-ylacetate (30.0 mg, 0.082 mmol) at room temperature and the reactionsolution was stirred at the same temperature for five hours. Thereaction solution was diluted with ethyl acetate and washed with asaturated sodium hydrogen carbonate aqueous solution, distilled waterand brine. After the organic layer was dried over anhydrous sodiumsulfate and the drying agent was filtered off, the organic layer wasconcentrated under reduced pressure. The obtained residue was purifiedby silica gel column chromatography (Kanto, commercial name Silica Gel60N, 40-100 μm; heptane:ethyl acetate=1:2) to obtain the title compound(33 mg) as a white solid.

400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.61 (q, J=8.0 Hz, 6H), 0.89 (d, J=6.8Hz, 3H), 0.97 (t, J=8.0 Hz, 9H), 1.20 (s, 3H), 1.34-1.50 (m, 3H),1.65-1.70 (m, 1H), 1.72 (d, J=1.2 Hz, 3H), 2.09 (s, 3H), 2.41 (dd,J=1.2, 14.0 Hz, 1H), 2.48 (dd, J=3.6, 14.0 Hz, 1H), 2.49-2.53 (m, 1H),3.80-3.90 (m, 1H), 4.98 (d, J=10.4 Hz, 1H), 5.07-5.09 (m, 1H), 5.15 (d,J=10.0 Hz, 1H), 5.24 (dd, J=1.6, 16.8 Hz, 1H), 5.58-5.70 (m, 2H), 6.14(d, J=10.8 Hz, 1H), 6.53 (dt, J=10.4, 16.8 Hz, 1H); 100 MHz ¹³C-NMR(CDCl₃) δ (ppm) 4.70, 6.80, 11.85, 16.47, 21.25, 24.80, 30.30, 35.57,40.48, 40.70, 70.16, 73.58 78.99, 82.23, 118.57, 125.09, 131.16, 131.99,133.58, 140.73, 168.67, 169.59; IR (neat)=3316, 2956, 2911, 2875, 1739,1459, 1370, 1240, 1170, 1089, 1020, 979, 908, 744 cm⁻¹; HRMSC₂₆H₄₄AgO₆Si (M+Ag⁺) Calcd: 587.1958, Found: 587.1915; [α]_(D) ²⁶+34.8(c 1.50, CHCl₃).

[Step D6]

Synthesis of Pladienolide D and its Analogues

(1) Synthesis of Pladienolide D

This reaction was performed with reference to the literature (Grubbs, R.H. “Handbook of Metathesis”, Wiley-VCH, 2003, v. 2, p 246-292).

(i)(2S,3S,4E,6S,7R,10R)-7,10-dihydroxy-3,7-dimethyl-2-[(1E)-1-methylbuta-1,3-dien-1-yl]-12-oxooxacyclododec-4-en-6-ylacetate (10.0 mg, 27.3 μmol) and(1R)-1,2-anhydro-3,5-dideoxy-1-[(1R,2S)-2-hydroxy-1-methylbutyl]-4-C-vinyl-D-erythro-pentitol(17.6 mg, 81.9 μmol) were dissolved in anhydrous dichloromethane (1 ml)under Ar atmosphere, followed by addition of the second generationHoveyda-Grubbs catalyst;[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(O-isopropoxyphenylmethylene)ruthenium(0.85 mg, 1.4 μmol). After the reaction solution was warmed to refluxfor one hour, it was cooled to room temperature and concentrated underreduced pressure. The obtained residue was purified by silica gel columnchromatography (Kanto, commercial name Silica Gel 60N, 40-100 μm;heptane:ethyl acetate=2.5:1→1:2→0:1) to obtain the title compound (9.5mg) as a white amorphous.

(ii)(2S,3S,4E,6S,7R,10R)-7,10-dihydroxy-3,7-dimethyl-2-[(1E)-1-methylbuta-1,3-dien-1-yl]-12-oxooxacyclododec-4-en-6-ylacetate (10.0 mg, 27.3 μmol) and(1R)-1,2-anhydro-3,5-dideoxy-1-[(1R,2S)-2-hydroxy-1-methylbutyl]-4-C-vinyl-D-erythro-pentitol(11.7 mg, 54.6 μmol) were dissolved in anhydrous dichloromethane (2.5ml) under Ar atmosphere, followed by addition of the second generationGrubbs catalyst;[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(phenylmethylene)-tricyclohexylphosphine)ruthenium(1.16 mg, 1.4 μmol). After the reaction solution was heated to refluxfor one hour, it was cooled to room temperature and concentrated underreduced pressure. The obtained residue was purified by silica gel columnchromatography (Kanto, commercial name Silica Gel 60N, 40-100 μm;heptane:ethyl acetate=2.5:1→1:2→0:1) to obtain the title compound (9.5mg) as a white amorphous.

600 MHz ¹H-NMR (CD₃OD) δ (ppm) 0.93 (d, J=6.8 Hz, 3H), 0.94 (d, J=7.0Hz, 3H), 0.98 (t, J=7.4 Hz, 3H), 1.23 (s, 3H), 1.30 (m, 1H), 1.38 (s,3H), 1.40 (m, 1H), 1.42 (m, 1H), 1.54 (m, 2H), 1.62 (m, 1H), 1.66 (m,1H), 1.70 (dd, J=6.3, 14.1 Hz, 1H), 1.82 (d, J=0.9 Hz, 3H), 1.91 (dd,J=5.5, 14.0 Hz, 1H), 2.10 (s, 3H), 2.57 (m, 2H), 2.62 (ddq, J=6.8, 9.8,10.6 Hz, 1H), 2.71 (dd, J=2.2, 7.9 Hz, 1H), 2.93 (ddd, J=2.2, 6.3, 6.3Hz, 1H), 3.57 (ddd, J=4.5, 4.5, 8.3 Hz, 1H), 3.83 (m, 1H), 5.09 (d,J=9.8 Hz, 1H), 5.11 (d, J=10.6 Hz, 1H), 5.61 (dd, J=9.9, 15.2 Hz, 1H),5.75 (dd, J=9.8, 15.2 Hz, 1H), 5.92 (d, J=15.3 Hz, 1H), 6.18 (d, J=11.0Hz, 1H), 6.57 (dd, J=11.0, 15.3 Hz, 1H); 150 MHz ¹³C-NMR (CD₃OD) δ (ppm)10.5, 10.8, 12.0, 16.9, 21.1, 24.2, 28.6, 28.8, 30.5, 37.5, 40.1, 41.8,42.6, 46.0, 56.0, 62.5, 70.4, 73.1, 74.1, 75.3, 80.3, 84.2, 123.7,127.1, 131.8, 133.8, 141.6, 143.2, 171.7, 172.2; IR (KBr)=3438, 2968,2939, 2879, 1733, 1720, 1458, 1371, 1244, 1176, 1021, 977, 832, 789,747, 612, 551, 474, 433, 420, 409 cm⁻¹; HRMS: C₃₀H₄₈NaO₉ (M+Na⁺), Calcd:575.3196, Found: 575.3168; [α]_(D) ²⁵−14.7 (c 1.10, MeOH).

(2) Synthesis of 3-[(triethylsilyl)oxy]-pladienolide D

This reaction was performed with reference to the literature (Grubbs, R.H. “Handbook of Metathesis”, Wiley-VCH, 2003, v. 2, p 246-292).

(2S,3S,4E,6S,7R,10R)-7-hydroxy-3,7-dimethyl-2-[(1E)-1-methylbuta-1,3-dien-1-yl]-12-oxo-10-[(triethylsilyl)oxy]oxacyclododec-4-en-6-ylacetate (8.60 mg, 17.9 μmol) and(1R)-1,2-anhydro-3,5-dideoxy-1-[(1R,2S)-2-hydroxy-1-methylbutyl]-4-C-vinyl-D-erythro-pentitol(8.67 mg, 40.5 μmol) were dissolved in anhydrous dichloromethane (1.8ml) under Ar atmosphere, followed by addition of the second generationGrubbs catalyst;[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(phenylmethylene)-tricyclohexylphospine)ruthenium(0.76 mg, 0.9 μmol). After the reaction solution was heated to refluxfor 1.5 hours, it was cooled to room temperature and concentrated underreduced pressure. The obtained residue was purified by silica gel columnchromatography (Kanto, commercial name Silica Gel 60N, 40-100 μm;heptane:ethyl acetate=3:1→2:1→1:1→0:1) to obtain the title compound(11.9 mg) as a colorless oil.

400 MHz ¹H-NMR (CD₃OD) δ (ppm) 0.68 (q, J=7.6 Hz, 6H), 0.93 (d, J=6.8Hz, 3H), 0.94 (d, J=7.2 Hz, 3H), 0.98 (t, J=7.2 Hz, 3H), 1.03 (t, J=7.6Hz, 9H), 1.21 (s, 3H), 1.26-1.31 (m, 1H), 1.32-1.36 (m, 2H), 1.38 (s,3H), 1.38-1.42 (m, 1H), 1.46-1.60 (m, 3H), 1.70 (dd, J=6.4, 14.0 Hz,1H), 1.80 (d, J=1.2 Hz, 3H), 1.91 (dd, J=5.6, 14.4 Hz, 1H), 2.10 (s,3H), 2.43 (dd, J=4.8, 14.0 Hz, 1H), 2.56 (dd, J=3.2, 14.0 Hz, 1H),2.58-2.66 (m, 1H), 2.71 (dd, J=2.4, 8.0 Hz, 1H), 2.94 (dt, J=2.4, 6.4Hz, 1H), 3.57 (dt, J=4.4, 8.0 Hz, 1H), 3.90-4.01 (m, 1H), 4.97 (d,J=10.8 Hz, 1H), 5.06 (d, J=9.6 Hz, 1H), 5.61 (dd, J=10.0, 15.2 Hz, 1H),5.75 (dd, J=9.6, 15.2 Hz, 1H), 5.91 (d, J=15.2 Hz, 1H), 6.18 (dd, J=1.2,11.2 Hz, 1H), 6.57 (dd, J=11.2, 15.2 Hz, 1H); HRMS: C₃₆H₆₂O₉NaSi(M+Na⁺), Calcd: 689.4061, Found: 689.4044.

(3) Synthesis of(3aS,4E,6S,7S,11R,13aR)-11-{[tert-butyl(dimethyl)silyl]oxy}-7-[(1E,3E)-5-hydroxy-1,5-dimethyl-7-phenylhepta-1,3-dien-1-yl]-6,13a-dimethyl-2-phenyl-3a,6,7,10,11,12,13,13a-octahydro-9H-[1,3]dioxolo[4,5-f]oxacyclododecin-9-one(Q15)

This reaction was performed with reference to the literature (Grubbs, R.H. “Handbook of Metathesis”, Wiley-VCH, 2003, v. 2, p 246-292).

(3aS,4E,6S,7S,11R,13aR)-11-{[tert-butyl(dimethyl)silyl]oxy}-6,13a-dimethyl-7-[(1E)-1-methylbuta-1,3-dien-1-yl]-2-phenyl-3a,6,7,10,11,12,13,13a-octahydro-9H-[1,3]dioxolo[4,5-f]oxacyclododecin-9-one(15.0 mg, 28.5 μmol) and 3-methyl-5-phenylpent-1-en-3-ol (15.1 mg, 85.5μmol) were dissolved in anhydrous dichloromethane (1.0 ml) under Aratmosphere, followed by addition of the second generation Grubbscatalyst;[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(phenylmethylene)-tricyclohexylphospine)ruthenium(2.0 mg, 2.4 μmol). The reaction solution was stirred at roomtemperature for 12 hours. Furthermore, the reaction solution was heatedto reflux for five hours. The reaction solution was cooled to roomtemperature and concentrated under reduced pressure. The obtainedresidue was purified by silica gel column chromatography (Merck,commercial name Silica Gel 60, 230-400 mesh; heptane:ethylacetate=6:1→3:1) to obtain the title compound (14.1 mg) as a colorlessoil. The title compound was determined to be a mixture of α benzylideneacetal and β benzylidene acetal in a ratio of 1:3 by ¹H-NMR.

Q15β: 400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.08 (s, 3H), 0.10 (s, 3H), 0.89(s, 9H), 0.93 (d, J=6.8 Hz, 3H), 1.24-1.28 (m, 1H), 1.35 (s, 3H), 1.37(s, 3H), 1.39-1.47 (m, 1H), 1.69 (t, J=11.6 Hz, 1H), 1.78 (s, 3H),1.80-1.94 (m, 2H), 1.99-2.04 (m, 1H), 2.32 (dd, J=6.0, 14.4 Hz, 1H),2.56-2.69 (m, 4H), 3.94-4.05 (m, 1H), 4.32 (d, J=10.0 Hz, 1H), 5.01 (d,J=10.8 Hz, 0.6H), 5.02 (d, J=10.8 Hz, 0.4H), 5.46 (dd, J=9.6, 15.2 Hz,1H), 5.86 (dd, J=15.2 Hz, 1H), 5.84-5.92 (m, 1H), 6.10 (s, 1H), 6.15 (d,J=10.8 Hz, 1H), 6.44-6.52 (m, 1H), 7.14-7.20 (m, 3H), 7.20-7.27 (m, 3H),7.30-7.39 (m, 2H), 7.47-7.52 (m, 2H); LRMS C₄₁H₅₈NaO₆Si (M+Na⁺) Calcd:697.39, Found: 697.44.

Reference Examples Synthesis of(3aS,4E,6S,7S,11R,13aR)-11-{[tert-butyl(dimethyl)silyl]oxy}-6,13a-dimethyl-7-[(1E)-1-methylbuta-1,3-dien-1-yl]-2-phenyl-3a,6,7,10,11,12,13,13a-octahydro-9H-[1,3]dioxolo[4,5-f]oxacyclododecin-9-one(Q10b)

(1) Synthesis of 3,21-bis{[tert-butyl(dimethyl)silyl]oxy}-pladienolide B(Q16)

Imidazole (1.66 g, 24.4 mmol) and tert-butyldimethylsilyl chloride (3.68g, 24.4 mmol) were added to a DMF (21.8 ml) solution of pladienolide B(2.18 g, 2.44 mmol, 60% purity) at room temperature and it was stirredat the same temperature for two hours. The reaction solution was dilutedwith ethyl acetate and washed with distilled water and brine. After theorganic layer was dried over anhydrous sodium sulfate and the dryingagent was filtered off, the organic layer was concentrated under reducedpressure. The obtained residue was purified by silica gel columnchromatography (Kanto, commercial name Silica Gel 60N, 40-100 μm;heptane:ethyl acetate=4:1) to obtain the title compound (1.68 g) as awhite amorphous.

LRMS C₄₂H₇₆NaO₈Si₂ (M+Na⁺) Calcd: 787.50, Found: 787.62.

(2) Synthesis of 3,21-bis{[tert-butyl(dimethyl)silyl]oxy}-pladienolide A(Q17)

Potassium carbonate (304 mg, 2.20 mmol) was added to a methanol (25.0ml) solution of 3,21-bis{[tert-butyl(dimethyl)silyl]oxy}-pladienolide B(1.68 g, 2.20 mmol) at room temperature. After the reaction solution wasstirred at room temperature for 1.5 hours, it was poured into brine,which was then extracted with ethyl acetate. After the organic layer wasdried over anhydrous sodium sulfate and the drying agent was filteredoff, the organic layer was concentrated under reduced pressure. Theobtained residue was purified by silica gel column chromatography(Kanto, commercial name Silica gel 60N, granular, neutral, 0.040mm-0.100 mm; heptane:ethyl acetate=3:1) to obtain the title compound(1.55 g) as a colorless oil.

400 MHz ¹H-NMR (CD₃OD) δ (ppm) 0.07 (s, 12H), 0.81 (t, J=7.6 Hz, 3H),0.85 (d, J=7.2 Hz, 6H), 0.91 (s, 18H), 1.08 (d, J=6.4 Hz, 3H), 1.25 (s,3H), 1.22-1.72 (m, 9H), 1.73 (s, 3H), 2.36 (dd, J=4.8, 13.6 Hz, 1H),2.40-2.50 (m, 1H), 2.50 (dd, J=3.2, 13.6 Hz, 1H), 2.50-2.60 (m, 1H),2.61 (dd, J=2.5, 8.4 Hz, 1H), 2.72 (dt, J=2.2, 6.0 Hz, 1H), 3.68 (d,J=9.6 Hz, 1H), 3.68-3.76 (m, 1H), 3.85-3.95 (m, 1H), 4.88 (d, J=12.4 Hz,1H), 5.38 (dd, J=10.0, 15.2 Hz, 1H), 5.62 (dd, J=8.4, 15.2 Hz, 1H) 5.71(dd, J=10.0, 15.2 Hz, 1H), 6.08 (brd, J=10.8 Hz, 1H), 6.32 (dd, J=10.8,15.2 Hz, 1H); LRMS C₄₀H₇₄NaO₇Si₂ (M+Na⁺) Calcd: 745.49, Found: 745.53.

(3) Synthesis of6,7-O-benzylidene-3,21-bis{[tert-butyl(dimethyl)silyl]oxy}-pladienolideA (Q18)

Benzaldehyde dimethylacetal (3.21 ml, 21.4 mmol) and pyridiniump-toluene sulfonate (53.8 mg, 0.214 mmol) were added to an anhydrousdichloromethane (16 ml) solution of3,21-bis{[tert-butyl(dimethyl)silyl]oxy}-pladienolide A (1.55 g, 2.14mmol). The reaction solution was stirred at room temperature for 28hours. The reaction solution was poured into a saturated sodium hydrogencarbonate aqueous solution, which was then extracted withdichloromethane. After the organic layer was washed with water and brinesequentially, it was dried over anhydrous magnesium sulfate. After thedrying agent was filtered off, the organic layer was concentrated underreduced pressure. The obtained residue was purified by silica gel columnchromatography (Kanto, commercial name Silica gel 60N, granular,neutral, 0.040 mm-0.100 mm; heptane:ethyl acetate=20:1→10:1) to obtainthe title compound (1.60 g) as a white amorphous. The title compound wasdetermined to be a mixture of α benzylidene acetal and β benzylideneacetal in a ratio of 1:3.2 by ¹H-NMR.

Compound Q18β: 400 MHz. ¹H-NMR (CD₃OD) δ (ppm) 0.08 (s, 6H), 0.11 (s,3H), 0.13 (s, 3H), 0.82 (t, J=7.6 Hz, 3H), 0.86 (t, J=7.2 Hz, 3H), 0.91(s, 18H), 0.92 (d, J=6.0 Hz, 3H), 1.08 (d, J=6.8 Hz, 3H), 1.21-1.31 (m,1H), 1.34 (s, 3H), 1.35-1.60 (m, 5H), 1.64-1.72 (m, 1H), 1.72-1.82 (m,1H), 1.76 (s, 3H), 1.90-2.00 (m, 1H), 2.25 (dd, J=10.0, 14.8 Hz, 1H),2.42-2.54 (m, 1H), 2.58-2.65 (m, 2H), 2.66-2.75 (m, 2H), 3.70-3.76 (m,1H), 4.00-4.10 (m, 1H), 4.37 (d, J=9.6 Hz, 1H), 4.96 (d, J=10.4 Hz, 1H),5.50 (dd, J=9.6, 15.2 Hz, 1H), 5.64 (dd, J=9.6, 15.2 Hz, 1H), 6.00 (dd,J=10.0, 14.8 Hz, 1H), 6.08 (s, 1H,), 6.09 (d, 9.6 Hz, 1H), 6.33 (dd,J=10.4, 14.8 Hz, 1H), 7.32-7.40 (m, 3H), 7.44-7.50 (m, 2H); LRMSC₄₇H₇₈NaO₇Si₂ (M+Na⁺) Calcd: 833.52, Found: 833.61.

(4) Synthesis of(3aS,4E,6S,7S,11R,13aR)-11-{[tert-butyl(dimethyl)silyl]oxy}-7-{(1E,5S)-6-[(2R,3R)-3-((1S,2S)-2-{[tert-butyl(dimethyl)silyl]oxy}-1-methylbutyl)oxiran-2-yl]-3,4-dihydroxy-1,5-dimethylhex-1-en-1-yl}-6,13a-dimethyl-2-phenyl-3a,6,7,10,11,12,13,13a-octahydro-9H-[1,3]dioxolo[4,5-f]oxacyclododecin-9-one(Q19)

6,7-O-benzylidene-3,21-bis{[tert-butyl(dimethyl)silyl]oxy}-pladienolideA (1.60 g, 1.97 mmol) was dissolved in THF (35 ml) and water (10 ml),followed by addition of osmium tetraoxide (1.25 ml, 0.197 mmol, 4%aqueous solution) and 4-methylmorpholine-4-oxide (0.4 ml, 2.17 mmol, 50%aqueous solution) at room temperature, and the mixture was stirred atthe same temperature for 24 hours. Subsequently sodium sulfite (0.498 g,3.94 mmol) was added to the reaction solution, and the mixture wasstirred at the same temperature for another hour. The reaction solutionwas diluted with ethyl acetate and washed with distilled water andbrine. After the organic layer was dried over anhydrous sodium sulfateand the drying agent was filtered off, the organic layer wasconcentrated under reduced pressure. The obtained residue was purifiedby silica gel column chromatography (Kanto, commercial name Silica Gel60N, 40-100 μm; heptane:ethyl acetate=2:1) to obtain the title compound(1.46 g) as a colorless oil.

LRMS C₄₇H₈₀NaO₉Si₂ (M+Na⁺) Calcd: 867.52, Found: 868.68.

(5) Synthesis of(5R)-4,5-anhydro-5-((1S,2S)-2-{[tert-butyl(dimethyl)silyl]oxy}-1-methylbutyl)-2,3-dideoxy-2-methyl-L-erythro-pentose(Q20) and(3aS,4E,6S,7S,13aR)-11-{[tert-butyl(dimethyl)silyl]oxy}-7-[(E)-2-formyl-1-methyleth-1-en-1-yl]-6,13a-dimethyl-2-phenyl-3a,6,7,10,11,12,13,13a-octahydro-9H-[1,3]dioxolo[4,5-f]oxacyclododecin-9-one(P18b)

(3aS,4E,6S,7S,11R,13aR)-11-{[tert-butyl(dimethyl)silyl]oxy}-7-{(1E,5S)-6-[(2R,3R)-3-((1S,2S)-2-{[tert-butyl(dimethyl)silyl]oxy}-1-methylbutyl)oxiran-2-yl]-3,4-dihydroxy-1,5-dimethylhex-1-en-1-yl}-6,13a-dimethyl-2-phenyl-3a,6,7,10,11,12,13,13a-octahydro-9H-[1,3]dioxolo[4,5-f]oxacyclododecin-9-one(1.46 g, 1.73 mmol) was dissolved in THF (30 ml) and water (7.5 ml),followed by addition of sodium metaperiodate (1.11 g, 5.19 mmol) at roomtemperature, and the mixture was stirred at the same temperature for 1.5hours. The reaction solution was diluted with ethyl acetate and washedwith distilled water and brine. After the organic layer was dried overanhydrous sodium sulfate and the drying agent was filtered off, theorganic layer was concentrated under reduced pressure. The obtainedresidue was purified by silica gel column chromatography (Kanto,commercial name Silica Gel 60N, 40-100 μm; heptane:ethylacetate=10:1→8:1→5:1) to obtain the title compound Q20 (0.485 g) as acolorless oil and to obtain the title compound P18b (0.803 g) as acolorless oil. The title compound P18b was determined to be a mixture ofα benzylidene acetal and β benzylidene acetal in a ratio of 1:3.2 by¹H-NMR. Compound Q20: 400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.06 (s, 6H), 0.81(t, J=7.6 Hz, 3H), 0.87 (t, J=6.8 Hz, 3H), 0.89 (s, 9H), 1.20 (dd,J=0.8, 7.6 Hz, 3H), 1.28-1.39 (m, 1H), 1.42-1.56 (m, 3H), 2.05 (ddd,J=4.4, 8.0, 13.6 Hz, 1H), 2.52-2.63 (m, 1H), 2.67 (dd, J=2.4, 8.0 Hz,1H), 2.72-2.80 (m, 1H), 3.67-3.76 (m, 1H), 9.67 (d, J=0.8 Hz, 1H); LRMSC₁₇H₃₅NaO₃Si (M+H⁺) Calcd: 315.24, Found: 315.06.

Compound Q18bβ: 400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.09 (s, 3H), 0.10 (s,3H), 0.89 (s, 9H), 0.98 (d, J=6.8 Hz, 3H), 1.35 (s, 3H), 1.34-1.50 (m,2H), 1.68-1.78 (m, 1H), 1.92-2.04 (m, 1H), 2.18 (brs, 3H), 2.37 (dd,J=9.6, 14.8 Hz, 1H), 2.60 (dd, J=4.0, 14.8 Hz, 1H), 2.60 (dd, J=4.0,14.8 Hz, 1H), 2.60-2.70 (m, 1H), 3.92-4.02 (m, 1H), 4.31 (d, J=10.0 Hz,1H), 4.94 (d, J=10.4 Hz, 1H), 5.46 (dd, J=9.6, 15.2 Hz, 1H), 5.92 (dd,J=9.6, 15.2 Hz, 1H), 6.07 (dd, J=1.6, 7.6 Hz, 1H), 6.11 (s, 1H),7.31-7.42 (m, 3H), 7.45-7.54 (m, 2H), 10.05 (d, J=7.6 Hz, 1H); LRMSC₃₀H₄₄NaO₆Si (M+Na⁺) Calcd: 551.28, Found: 551.29.

(6) Synthesis of(3aS,4E,6S,7S,11R,13aR)-11-{[tert-butyl(dimethyl)silyl]oxy}-6,13a-dimethyl-7-[(1E)-1-methylbuta-1,3-dien-1-yl]-2-phenyl-3a,6,7,10,11,12,13,13a-octahydro-9H-[1,3]dioxolo[4,5-f]oxacyclododecin-9-one(Q10b)

Potassium tert-butoxide (38.2 mg, 0.341 mmol) was added to an anhydrousTHF (3 ml) solution of methyltriphenylphosphonium iodide (100 mg, 0.247mmol) at room temperature, and it was stirred at room temperature for 30minutes. Subsequently, a THF (3 ml) solution of(3aS,4E,6S,7S,13aR)-11-{[tert-butyl(dimethyl)silyl]oxy}-7-[(E)-2-formyl-1-methyletha-1-en-1-yl]-6,13a-dimethyl-2-phenyl-3a,6,7,10,11,12,13,13a-octahydro-9H-[1,3]dioxolo[4,5-f]oxacyclododecin-9-one(150 mg, 0.284 mmol) was added dropwise to this reaction solution atroom temperature and the reaction solution was stirred for one hour. Thereaction solution was diluted with ethyl acetate and washed withdistilled water and brine. After the organic layer was dried overanhydrous sodium sulfate and the drying agent was filtered off, theorganic layer was concentrated under reduced pressure. The obtainedresidue was purified by silica gel column chromatography (Kanto,commercial name Silica Gel 60N, 40-100 μm; heptane:ethyl acetate=20:1)to obtain the title compound (44 mg) as a white solid. The titlecompound was determined to be a mixture of α benzylidene acetal and βbenzylidene acetal in a ratio of 1:3.2 by ¹H-NMR.

Q10bβ: 400 MHz ¹H-NMR (CDCl₃) δ (ppm) 0.08 (s, 3H), 0.10 (s, 3H), 0.89(s, 9H), 0.92 (d, J=6.8 Hz, 3H), 1.28-1.50 (m, 2H), 1.34 (s, 3H),1.65-1.74 (m, 1H), 1.75 (brs, 3H), 1.98-2.08 (m, 1H), 2.32 (dd, J=10.0,14.4 Hz, 1H), 2.58 (dd, J=4.0, 14.4 Hz, 1H), 2.58-2.70 (m, 1H),3.92-4.02 (m, 1H), 4.32 (d, J=10.0 Hz, 1H), 5.00 (d, J=10.8 Hz, 1H),5.18 (d, 10.0 Hz, 1H), 5.27 (d, J=16.8 Hz, 1H), 5.46 (dd, J=9.6, 15.2Hz, 1H), 5.89 (dd, J=10.0, 15.2 Hz, 1H), 6.09 (s, 1H), 6.14 (d, J=10.8Hz, 1H), 6.55 (dt, J=10.8, 16.8 Hz, 1H), 7.28-7.41 (m, 3H), 7.44-7.53(m, 2H); LRMS C₃₁H₄₆NaO₅Si (M+Na⁺) Calcd: 549.30, Found: 549.26.

(6-1) Alternative Synthesis of(3aS,4E,6S,7S,11R,13aR)-11-{[tert-butyl(dimethyl)silyl]oxy}-6,13a-dimethyl-7-[(1E)-1-methylbuta-1,3-dien-1-yl]-2-phenyl-3a,6,7,10,11,12,13,13a-octahydro-9H-[1,3]dioxolo[4,5-f]oxacyclododecin-9-one(Q10b)

The reaction was performed with reference to the literature (Ager, D.J., Synthesis, 1984, 384-398. Hudrlik, P. F.; Peterson, D., J. Am. Chem.Soc., 1975, 97, 1464-1468.).

Magnesium trimethylsilylmethylchloride 1.5M diethyl ether solution (1.33ml, 2.0 mmol) was added to a diethyl ether solution (20 ml) of(2S,3aS,4E,6S,7S,13aR)-11-{[tert-butyl(dimethyl)silyl]oxy}-7-[(E)-2-formyl-1-methyleth-1-en-1-yl]-6,13a-dimethyl-2-phenyl-3a,6,7,10,11,12,13,13a-octahydro-9H-[1,3]dioxolo[4,5-f]oxacyclododecin-9-one(510 mg, 0.965 mmol) and stirred at the same temperature for 30 minutes.The reaction solution was diluted with diethyl ether, followed byaddition of water and sequentially washed with ammonium chloride aqueoussolution and brine. The organic layer was dried over anhydrous magnesiumsulfate, the drying agent was filtered off and the organic layer wasconcentrated under reduced pressure.

The obtained crude product was dissolved in THF (15 ml), followed byaddition of sulfuric acid (6 drops) at room temperature and the solutionwas stirred for 1 hour at the same temperature. The reaction solutionwas diluted with ethyl acetate and washed with water and brine. Theorganic layer was dried over anhydrous magnesium sulfate, the dryingagent was filtered off and the organic layer was concentrated underreduced pressure. The obtained residue was purified by silica gel columnchromatography (Merck, commercial name Silica Gel 60, 230-400 mesh;hexane:ethyl acetate=30:1) to obtain the title compound (312 mg) ascolorless oil. ¹H-NMR data of title compound by this alternative methodare completely identical to those of the title compound obtained by theabove (6).

Reference Example 2

Synthesis of(2S,3S,4E,6S,7R,10R)-10-{[tert-butyl(dimethyl)silyl]oxy}-2-{(1E,5S)-6-[(2R,3R)-3-((1S,2S)-2-{tert-butyl(dimethyl)silyl}oxy)-1-methylbutyl]oxiran-2-yl}-3,4-dihydroxy-1,5-dimethylhexa-1-en-1-yl}-7-hydroxy-3,7-dimethyl-12-oxooxacyclododeca-4-en-6-ylacetate (R6)

3,21-bis{[tert-butyl(dimethyl)silyl]oxy}-pladienolide B (60 mg, 0.078mmol) was dissolved in THF (1 ml) and water (1 ml), followed by additionof osmium tetraoxide (0.199 ml, 0.016 mmol, 2% aqueous solution) and4-methylmorpholine-4-oxide (18 μml, 0.078 mmol, 50% aqueous solution) atroom temperature. The solution was stirred for 24 hours at the sametemperature. Then sodium sulfite (19.8 mg, 0.156 mmol) was added to thereaction solution and further stirred for 1 hour at the sametemperature. The reaction solution was diluted by ethyl acetate andwashed with the distilled water and brine. The organic layer was driedover anhydrous sodium sulfate, the drying agent was filtered off and theorganic layer was concentrated under reduced pressure. The obtainedresidue was purified by silica gel column chromotography (Kanto,commercial name Silica Gel 60N, 40-100 μm; heptane:ethyl acetate=1:1) toobtain the title compound (67.6 mg) as colorless oil. The title compoundwas determined as a mixture of diastereomers (1:3).

400 MHz ¹H-NMR (CD₃OD) δ (ppm) 0.11 (s, 12H), 0.85-0.89 (m, 3H),0.92-0.98 (m, 6H), 0.95 (s, 9H), 0.96 (s, 9H), 0.99 (d, J=6.0 Hz,0.75H), 1.11 (d, J=6.0 Hz, 2.25H), 1.21 (s, 3H), 1.24-1.74 (m, 9.25H),1.73 (s, 2.25H), 1.79 (s, 0.75H), 1.84-1.95 (m, 0.75H), 2.10 (s, 3H),2.36-2.45 (m, 1H), 2.51-2.63 (m, 2H), 2.61 (dd, J=2.0, 8.0 Hz, 0.75H),2.69 (dd, J=2.0, 8.0 Hz, 0.25H), 2.72-2.84 (m, 1H), 3.28 (dd, J=5.6, 5.6Hz, 0.75H), 3.45 (dd, J=2.8, 7.2 Hz, 0.25H), 3.74-3.82 (m, 1H),3.90-4.00 (m, 1H), 4.26-4.39 (m, 1H), 4.87-4.93 (1H, covered withCD₃OD), 4.96 (d, J=9.6 Hz, 0.25H), 5.06 (d, J=9.6 Hz, 0.75H), 5.54-5.68(m, 2H), 5.75 (dd, J=9.6 Hz, 15.2 Hz, 1H); LRMS C₄₂H₇₈NaO₁₀Si₂ (M+Na⁺)Calcd: 821.50, Found: 821.26.

Reference Example 3

Synthesis of(5R)-4,5-anhydro-5-((1S,2S)-2-{[tert-butyl(dimethyl)silyl]oxy}-1-methylbutyl)-2,3-dideoxy-2-methyl-L-erythro-pentose(Q20) and(2S,3S,4E,6S,7R,10R)-10-{[tert-butyl(dimethyl)silyl]oxy}-7-hydroxy-3,7-dimethyl-2-[(1E)-1-methyl-3-oxoprop-1-en-1-yl]-12-oxooxacyclododec-4-en-6-ylacetate (R4)

(2S,3S,4E,6S,7R,10R)-10-{[tert-butyl(dimethyl)silyl]oxy}-2-{(1E,5S)-6-[(2R,3R)-3-((1S,2S)-2-{[tert-butyl(dimethyl)silyl]oxy}-1-methylbutyl)oxiran-2-yl]-3,4-dihydroxy-1,5-dimethylhex-1-en-1-yl}-7-hydroxy-3,7-dimethyl-12-oxooxacyclododeca-4-en-6yl acetate (67.6 mg 0.0847 mmol) was dissolved in THF (1.5 ml) and water(1.5 ml), followed by addition of sodium metaperiodate (181 mg, 0.847mmol) at room temperature. The reaction solution was stirred for 1 hourat the same temperature. The reaction solution was diluted by distilledwater and brine. After the organic layer was dried over anhydrous sodiumsulfate, the drying agent was filtered off and the organic layer wasconcentrated under reduced pressure. The obtained residue was purifiedby silica gel column chromotography (Kanto, commercial name Silica Gel60N, 40-100 μm; heptane:ethyl acetate=20:1→2:1) to obtain the titlecompound R4 (19.3 mg) as colorless oil and the title compound Q20 (36.0mg) as colorless oil. ¹H-NMR data of title compound are completelyidentical to those of the compound Q20 obtained by Reference Example 1(5).

R4: 400 MHz ¹H-NMR (CD₃OD) δ (ppm) 0.12 (s, 3H), 0.14 (m, 3H), 0.95 (s,9H), 1.00 (d, J=6.8 Hz, 3H), 1.24 (s, 3H), 1.23-1.84 (m, 4H), 2.10 (s,3H), 2.22 (d, J=1.2 Hz, 3H), 2.44 (dd, J=4.8, 13.6 Hz, 1H), 2.55-2.66(m, 2H), 3.93-4.03 (m, 1H), 4.93 (d, J=10.8 Hz, 1H), 5.06 (d, J=9.6,1H), 5.64 (dd, J=10.0, 15.2 Hz, 1H), 5.80 (dd, J=9.6, 15.2 Hz, 1H), 6.08(brd, J=8.0 Hz, 1H), 10.06 (d, J=8.0 Hz, 1H); LRMS C₂₅ClH₄₂OSi (M+Cl⁻)Calcd: 517.24, Found: 7.09.

Reference Example 4

Synthesis oftert-butyl[((1S,2S)-1-ethyl-2{(2R,3R)-3-[(2S)-2-methylbut-3-en-1-yl]oxiran-2-yl}propyl)oxy]dimethylsilane(R5)

This reaction was performed with reference to the literature (Matusbara,S.; Sugihara, M,; Utimoto, K., Synlett, 1998, 97, 313-315.).

Boron trifluoride dimethyl ether complex was added to THF solution (6ml) of 10% wt of tetrahydrofuran suspension (4.56 g, 2.0 mmol) of Nystedreagent ([cyclo-dibromodi-μ-methylene(μ-tetrahydrofuran)trizinc]) at 0°C. and stirred for 5 minutes at the same temperature. THF solution (2ml) of((5R)-4,5-anhydro-5-((1S,2S)-2-{[tert-butyl(dimethyl)silyl]oxy}-1-methylbutyl)-2,3-dideoxy-2-methyl-L-erythro-pentose(315 mg, 0.01 mmol) was added to the reaction solution. After thesolution was warmed to room temperature, it was stirred for 3 hours. 1Mof Hydrochloric acid (3 ml) was added to the reaction solution and itwas diluted with hexane, sequentially washed with distilled water andbrine. It was dried over anhydrous magnesium sulfate, the drying agentwas filtered off and concentrated under reduced pressure. The obtainedresidue was purified by silica gel column chromotography (Kanto,commercial name Silica Gel 60N, 40-100 μm; heptane:ethylacetate=50:1→5:1) to obtain the title compound (9.77 mg) as colorlessoil. R4: 400 MHz ¹H-NMR (CD₃OD) δ (ppm) 0.13 (s, 6H), 0.88 (d, J=7.6 Hz,3H), 0.94 (d, J=7.6 Hz, 3H), 0.96 (s, 9H), 1.10 (d, J=6.8 Hz, 3H),1.30-1.42 (m, 1H), 1.48-1.64 (m, 4H), 2.34-2.50 (m, 1H), 2.72 (dd,J=2.4, 8.0 Hz, 1H), 2.75-2.85 (m, 1H), 3.69-3.82 (m, 1H) 5.01 (brd,J=10.8 Hz, 1H), 5.08 (brd, J=17.2 Hz, 1H) 5.72-5.86 (m, 1H); LRMSC₁₈H₃₆NaO₂Si (M+Na⁺) Calcd: 335.24, Found: 335.05.

Reference Example 5

Synthesis of(2S,3S,4E,6S,7R,10R)-7-hydroxy-3,7-dimethyl-2-[(1E)-1-methylbuta-1,3-dien-1-yl]-12-oxo-10-{[tert-butyl(dimethyl)silyl]oxy}oxacyclododec-4-en-6-ylacetate (R6)

This reaction was performed with reference to the literature (Pine, S.H.; Zahler, R.; Evans, D. A. Grubbs, R. H., J. Am. Chem. Soc., 1980,3270-3272.).

THF (0.8 ml) of(2S,3S,4E,6S,7R,10R)-10-{tert-butyl(dimethyl)silyl]oxy}-7-hydroxy-3,7-dimethyl-2-[(1E)-1-methyl-3-oxoprop-1-en-1-yl]-12-oxooxacyclododec-4-en-6-ylacetate (86.9 mg, 0.18 mmol) and toluene (6.1 ml) was dissolved as amixed solvent and it was cooled to −40° C. Pyridine (0.15 ml, 1.89 mmol)and 0.5 M toluene solution of Tebbe reagent(chlorobis(cyclopentadienyl)-(dimethylaluminium)-(methylenetitanium)were added to it and the reaction solution was stirred for 30 minutes atthe same temperature. The reaction solution was warmed to −20° C. andstirred for further 30 minutes. Distilled water was added to thereaction solution and it was diluted with ethyl acetate and washedsequentially with distilled water and brine. After the reaction solutionwas dried over anhydrous sodium sulfate, the drying agent was filteredoff and was concentrated under reduced pressure. The obtained residuewas purified by silica gel column chromatography (Merck, commercialname; Silica Gel 60, 230-400 mesh; hexane; diethyl ether=4:1) to obtainthe title compound (41 mg) as a colorless amorphous.

400 MHz ¹H-NMR (CDCL₃) δ (ppm) 0.06 (s, 3H), 0.07 (s, 3H), 0.89 (d,J=6.4 Hz, 3H), 0.97 (s, 9H), 1.21 (s, 3H), 1.28-1.50 (m, 3H), 1.62-1.70(m, 1H), 1.72 (brs, 3H), 2.10 (s, 3H), 2.39 (dd, J=4.4, 14.0 Hz, 1H),2.46 (dd, J=3.6, 14.0 Hz, 1H), 2.44-2.55 (m, 1H), 3.80-3.90 (m, 1H),4.97 (d, J=10.8 Hz, 1H), 5.07-5.09 (m, 1H), 5.15 (dd, J=1.6, 10.0 Hz,1H), 5.24 (dd, J=1.6, 16.8 Hz, 1H), 5.56-5.72 (m, 2H), 6.14 (d, J=10.8Hz, 1H), 6.53 (dt, J=10.0, 16.8 Hz, 1H); LRMS C₂₆H₄₄NaO₆Si (M+Na⁺)Calcd: 503.28, Found: 503.21.

Reference Example 6

Synthesis of4,5-anhydro-3,6,8,9-tetradeoxy-6-methyl-2-C-methyl-2,7-bis-O-(triethylsilyl)-L-glycero-D-gluco-nonose(R8)

3,16,21-tris-[(triethylsilyl)oxy]-pladienolide D (50 mg, 55.9 μl;Example 46 of WO 03/099813) was dissolved in dichloromethane. Thereaction solution was cooled to −78° C. Ozone (flow rate 2 l/min,voltage 90 V) was blown into the reaction solution for 10 minutes duringstirring. After dimethyl sulfide (10,4 mg, 0.168 mmol) was added, thereaction solution was warmed to room temperature, stirred for 1 hour andwas concentrated under reduced pressure. The obtained residue waspurified by silica gel column chromatography (Kanto, commercial nameSilica Gel 60N, granular, neutral, 0.040 mm-0.100 mm; n-heptane:ethylacetate=30:1) to obtain the title compound (18.5 mg) as colorless oil.

400 MHz ¹H-NMR (CD₃OD) δ (ppm) 0.64-0.76 (m, 12H), 0.88 (t, J=7.6 Hz,3H), 0.94 (d, J=7.6 Hz, 3H), 0.99 (t, J=7.6 Hz, 9H), 1.00 (t, J=7.6 Hz,9H), 1.28-1.39 (m, 1H), 1.42 (s, 3H), 1.52-1.63 (m, 2H), 1.81-1.94 (m,2H), 2.70 (dd, J=2.2, 8.0 Hz, 1H), 2.92 (dt, J=2.2, 8.0 Hz, 1H), 3.80(dt, J=3.2, 6.0 Hz, 1H), 9.61 (s, 1H).

Reference Example 7

Synthesis of5,6-anhydro-1,2,4,7-tetradeoxy-4-methyl-8-C-methyl-3,8-bis-O-triethylsilyl)-L-glycero-D-galacto-nonitol(R9),(1R)-1,2-anhydro-3,5-dideoxy-4-C-[(1E)-3-hydroxyprop-1-en-1-yl]-1-{(1S,2S)-1-methyl-2-[(triethylsilyl)oxy]butyl}-4-O-(triethylsilyl)-D-erythro-pentitol(R10),(2S,3S,4E,6S,7R,10R)-7-hydroxy-2-[(1E)-3-hydroxy-1-methylprop-1-en-1-yl]-3,7-dimethyl-12-oxo-10-[(triethysilyl)oxy]oxacyclododec-4-en-6-ylacetate (R11) and(2S,3S,4E,6S,7R,10R)-7-hydroxy-2-[(hydroxyethyl)-3,7-dimethyl-12-oxo-10-[(triethylsilyl)oxy]oxacyclododec-4-en-6-ylacetate (R12)

3,16,21-tris-[(triethylsilyl)oxy]-pladienolide D (21 mg, 0.235 mmol) wasdissolved in a mixture solvent of dichloromethane (25 ml) and methanol(25 ml). This reaction solution was cooled to −78° C. and Ozone (flowrate 2 l/min, voltage 90 V) was blown into the reaction solution for 3minutes during stirring. After adding sodium borohydride (88.9 mg, 2.35mmol), the reaction solution was warmed to 0° C. and stirred for 1.5hours. The reaction solution was diluted by ethyl acetate and washedwith brine. The organic layer was dried over anhydrous sodium sulfate,the drying agent was filtered off and the organic layer was concentratedunder reduced pressure. The obtained residue was purified by silica gelcolumn chromatography (Merck, commercial name Silica Gel 60, 230-400mesh; hexane:ethyl acetate=10:1→3:2→1:2). The title compound R9 (77.3mg) was obtained as colorless oil. The title compound R10 (17.3 mg) wasobtained as colorless oil. The title compound R11 (38.5 mg) was obtainedas colorless oil. The title compound R12 (49.1 mg) was obtained ascolorless oil.

R9: 400 MHz ¹H-NMR (CD₃OD) δ (ppm) 0.64-0.76 (m, 12H), 0.89 (t, J=7.2Hz, 3H), 0.98 (d, J=7.6 Hz, 3H), 0.95 (t, J=8.0 Hz, 9H), 1.00 (t, J=8.0Hz, 9H), 1.26 (s, 3H), 1.28-1.40 (m, 1H), 1.52-1.65 (m, 2H), 1.68 (dd,J=6.4, 14.4 Hz, 1H), 1.77 (dd, J=5.2, 14.4 Hz, 1H), 2.71 (dd, J=2.4, 8.0Hz, 1H), 2.95-3.02 (m, 1H), 3.48 (d, J=9.6 Hz, 1H), 3.55 (d, J=9.6 Hz,1H), 3.79 (dt, J=3.2, 6.4 Hz, 1H), LRMS C₂₃H₅₀NaO₄Si₂ (M+Na⁺) Calcd:469.31, Found: 469.16

R10: 400 MHz ¹H-NMR (CD₃OD) δ (ppm) 0.69 (q, J=8.0 Hz, 6H), 0.70 (q,J=8.0 Hz, 6H), 0.89 (t, J=7.6 Hz, 3H), 0.96 (d, J=7.2 Hz, 3H), 1.02 (t,J=8.0 Hz, 9H), 1.03 (t, J=8.0 Hz, 9H), 1.28-1.40 (m, 1H), 1.46 (s, 3H),1.52-1.66 (m, 1H), 1.70-1.84 (m, 1H), 2.69 (dd, J=2.4, 8.4 Hz, 1H), 2.95(dt, J=2.4, 5.6 Hz, 1H), 3.81 (dt, J=3.2, 6.4 Hz, 1H), 4.12 (d, J=4.0Hz, 2H), 5.78-5.88 (m, 2H), LRMS C₂₅H₅₂NaO₄Si₂ (M+Na⁺) Calcd: 495.33,Found: 495.31.

R11: LRMS C₂₅H₄₄NaO₇Si (M+Na⁺) Calcd: 507.28, Found: 507.27.

R12: 400 MHz ¹H-NMR (CD₃OD) δ (ppm) 0.66 (q, J=8.0 Hz, 6H), 1.02 (t,J=8.0 Hz, 9H), 1.11 (d, J=6.8 Hz, 3H), 1.16 (d, J=6.4 Hz, 3H), 1.24 (s,3H), 1.34-1.72 (m, 4H), 2.13 (s, 3H), 2.45-2.68 (m, 2H), 2.73-2.85 (m,1H), 3.85-3.95 (m, 1H), 3.98-4.12 (m, 1H), 4.77 (d, J=10.4 Hz, 1H), 5.13(d, J=9.2 Hz, 1H), 5.64-5.80 (m, 2H); LRMS C₂₃H₄₂NaO₇Si (M+Na⁺) Calcd:481.26, Found: 481.22.

1. A process for producing a compound represented by Formula (1):

wherein P¹ and R¹ are the same as defined below, characterized bycomprising reacting a compound represented by Formula (2):

wherein P¹ means a hydrogen atom or a protecting group for hydroxygroup; and R¹ means a hydrogen atom or a hydroxy group, with anacetylating agent in the presence of a base.
 2. The production processaccording to claim 1, wherein the acetylating agent is acetic anhydride.3. The production process according to claim 1 or 2, wherein the base isa combination of triethylamine with 4-dimethylaminopyridine.
 4. Theproduction process according to any one of claims 1 to 3, wherein thecompound represented by Formula (2) in which P¹ and R¹ are hydrogenatoms is obtained by removing protecting groups for hydroxy group at 3-and 21-positions of a compound represented by Formula (6):

wherein P² means a protecting group for hydroxy group; P³ means aprotecting group for hydroxy group; and R² means a phenyl group whichmay have a substituent, to obtain a compound represented by Formula (5):

wherein R² is the same as defined above, subsequently protecting hydroxygroups at the 3- and 21-positions of Compound (5) to obtain a compoundrepresented by Formula (4):

wherein P⁴ and P⁵ mean protecting groups for hydroxy group; and R² isthe same as defined above, subsequently removing protecting groups forhydroxy group at 6- and 7-positions of Compound (4) >—R² to obtain acompound represented by Formula (3):

wherein P⁴ and P⁵ are the same as defined above, and subsequentlyremoving protecting groups for hydroxy group at 3- and 21-positions ofCompound (3).
 5. A process for producing a compound represented byFormula (6):

wherein P², P³ and R² are the same as defined below, characterized bycomprising reacting a compound represented by Formula (7):

wherein P³ means a protecting group for hydroxy group; and Het means a1-phenyl-1H-tetrazol-5-yl group, with a compound represented by Formula(8):

wherein P² means a protecting group for hydroxy group; and R² means aphenyl group which may have substituents, in the presence of a base. 6.The production process according to claim 5, wherein the base ispotassium bis(trimethylsilyl)amide.
 7. A process for producing acompound represented by Formula (9):

wherein P², P⁶ and R² are the same as defined below, characterized bycomprising closing a ring in a compound represented by Formula (10)

wherein P² means a protecting group for hydroxy group; R² means a phenylgroup which may have substituents; and P⁶ means a protecting group forhydroxy group, in the presence of a catalyst.
 8. The production processaccording to claim 7, wherein the catalyst is[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(O-isopropoxyphenylmethylene)ruthenium.9. A process for producing a compound represented by Formula (11):

wherein P¹, P⁷, P⁸, P⁹ and R¹ are the same as defined below,characterized by comprising reacting a compound represented by Formula(12):

wherein P⁷ means a hydrogen atom or a protecting group for hydroxygroup; and R¹ means a hydrogen atom or a hydroxy group, with a compoundrepresented by Formula (13):

wherein P¹ means a hydrogen atom or a protecting group for hydroxygroup; P⁸ means a hydrogen atom, an acetyl group or a protecting groupfor hydroxy group; P⁹ means a hydrogen atom or a protecting group forhydroxy group; or P⁸ and P⁹ may form together a group represented by aformula:

wherein R⁵ means a phenyl group which may have substituents, in thepresence of a catalyst.
 10. The production process according to claim 9,wherein the catalyst is[1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene]dichloro(O-isopropoxyphenylmethylene)rutheniumortricyclohexylphosphine[1,3-bis(2,4,6-trimethylphenyl)-4,5-dihydroimidazol-2-ylidene][benzylidene]ruthenium(IV) dichloride.
 11. A compound represented by Formula (3-1):

wherein P⁴¹ and P⁵¹ may be the same or different and mean a hydrogenatom or a protecting group for hydroxy group, provided that both of P⁴¹and P⁵¹ are or either P⁴¹ or P⁵¹ is a protecting group for hydroxygroup, or a salt thereof.
 12. A compound represented by Formula(4,5,6-1)

wherein P²⁴¹ means a hydrogen atom or a protecting group for hydroxygroup; P³⁵¹ means a hydrogen atom or a protecting group for hydroxygroup; and R²¹ means a phenyl group which may have substituents, or asalt thereof.
 13. A compound represented by Formula (7-1):

wherein P³¹ means a hydrogen atom or a protecting group for hydroxygroup; and Het means a 1-phenyl-1H-tetrazol-5-yl group, or a saltthereof.
 14. A compound represented by Formula (8-1)

wherein P²¹ means a hydrogen atom or a protecting group for hydroxygroup; R²¹ means a phenyl group which may have substituents; and R⁴¹means a formyl group or a group represented by —CH₂OP⁶¹, wherein P⁶¹means a hydrogen atom or a protecting group for hydroxy group, or a saltthereof.
 15. A compound represented by Formula (10-1):

wherein P²¹ means a hydrogen atom or a protecting group for hydroxygroup; P⁶¹ means a hydrogen atom or a protecting group for hydroxygroup; and R²¹ means a phenyl group which may have substituents, or asalt thereof.
 16. A compound represented by Formula (12-1)

wherein P⁷¹ means a hydrogen atom or a protecting group for hydroxygroup; and R¹¹ means a hydrogen atom or a hydroxy group, or a saltthereof.
 17. A compound represented by Formula (13-1):

wherein P¹¹ means a hydrogen atom or a protecting group for hydroxygroup; P⁸¹ means a hydrogen atom, an acetyl group or a protecting groupfor hydroxy group; and P⁹¹ means a hydrogen atom or a protecting groupfor hydroxy group; or P⁸¹ and P⁹¹ may form together a group representedby a formula:

wherein R⁵¹ means a phenyl group which may have substituents, or a saltthereof.
 18. A process for producing a compound represented by Formula(11′):

wherein P¹, P⁸, P⁹, R⁶, R⁷ and R⁸ are the same as defined below,characterized by comprising reacting a compound represented by Formula(12′):

wherein R⁶ means a hydrogen atom or a linear C₁₋₁₀ alkyl group which mayhave a substituent; R⁷ means a hydrogen atom or a methyl group; and R⁸means a hydrogen atom or a hydroxy group, with a compound represented byFormula (13):

wherein P¹ means a hydrogen atom or a protecting group for hydroxygroup; P⁸ means a hydrogen atom, an acetyl group or a protecting groupfor hydroxy group; and P⁹ means a hydrogen atom or a protecting groupfor hydroxy group; or P⁸ and P⁹ may form together a group represented bya formula:

wherein R⁵ means a phenyl group which may have substituents, in thepresence of a catalyst.
 19. A compound represented by Formula (14):

wherein P⁷¹¹ means a hydrogen atom or a protecting group for hydroxygroup; and R¹¹¹ means a hydrogen atom or a hydroxy group; and R^(A)means a formyl group or a group represented by —CH₂OH, or a saltthereof.
 20. A compound represented by Formula (16):

wherein P²¹¹ means a hydrogen atom or a protecting group for hydroxygroup; and P⁸¹¹ means a hydrogen atom, an acetyl group or a protectinggroup for hydroxy group; and P⁹¹¹ means a hydrogen atom or a protectinggroup for hydroxy group; and R⁴¹¹ means a formyl group or a grouprepresented by —CH₂OP⁶¹¹ (wherein P⁶¹¹ means a hydrogen atom or aprotecting group for hydroxy group); and provided that a compoundrepresented by the following formula is excluded, or a salt thereof.